VETERINARY DIAGNOSTIC TEST KITS & REAGENTS

Transcription

1 2006 VMRD VETERINARY DIAGNOSTIC TEST KITS & REAGENTS

2 For the 26 years I have been privileged to be part of VMRD, God has graciously blessed, prospered and protected us. The words of Jesus Christ, Do unto others as you would have them do unto you, are the basis of my policy toward our customers and employees. Consistent with this policy is our goal to produce the best possible products, backed by sound technical service. Should we fail to achieve this goal, please feel free to contact me personally. D. Scott Adams, DVM, PhD, President and CEO

3 VMRD PRODUCT CATALOG DIAGNOSTIC TEST KITS FA REAGENTS ANTIBODIES IMMUNOLOGY REAGENTS TEST KITS FA REAGENTS ANTIBODIES IMMUNOLOGY

4 NEW PRODUCTS NEW TEST KITS Equine Infectious Anemia Virus Antibody Test Kit, ELISA...15 ELISAWare plate-reading software...5 NEW FA REAGENTS Anaplasma phagocytophila negative control Anaplasma phagocytophila positive control Babesia bigemina negative control Babesia bigemina positive control Babesia bovis negative control Babesia bovis positive control Bartonella henselae IgG positive control Bartonella henselae IgM positive control Bartonella henselae 12-well IFA substrate slide Bovine Parainfluenza Type 3 negative control Bovine Parainfluenza Type 3 positive control Canine Adenovirus negative control Canine Adenovirus positive control Affinity Purified FITC anti-canine IgG conjugate Transmissible Gastroenteritis Virus negative control Transmissible Gastroenteritis Virus positive control Vesicular Stomatitis Virus direct FA conjugate NEW ANTIBODIES Anti-Bovine Leukemia Virus antisera...31 Anti-Canine Parainfluenza Virus antisera...31 IL-A116 CD45R0 MoAb IgG3 isotype

5 TEST KITS VMRD Presents ELISAWare TM VMRD is pleased to announce the arrival of ELISAWare TM, the microplate- reading software that supports all VMRD ELISA test kits. The software will retrieve data from a microplate absorbance reader, display the data, validate the assay, calculate qualitative results, display the results, store sample identifications and results, and generate reports. Report options include a detailed analytical report for internal laboratory use or a client report displaying only the information relevant to a particular client. Exporting OD values to Microsoft Excel is as easy as clicking your mouse! Currently, ELISAWare TM supports microplate readers from four major manufacturers. If your reader is not supported, please contact VMRD by phone, fax, or and we will do our best to add your driver to ELISAWare. TM Don t see your reader? Let us know! We may be able to add a driver for your reader to ELISAWare! ELISAWare TM will validate and calculate results for all of VMRD s test kits. It can retrieve ODs from a plate reader for any given ELISA but will only validate and calculate results for VMRD s assays. As we bring new kits to market we will offer upgrades that keep your software current with all of our newest ELISAs. ELISAWare TM displays its reports in your Internet browser, providing multiple options for displaying, exporting, and analyzing ELISA results. ELISAWare TM was developed to be user-friendly, and VMRD is committed to offering professional and courteous technical support. We developed this software with our customers in mind, and we want it to work your way. If you would like to see a change in ELISAWare TM please let us know! We need your input to make ELISAWare TM the perfect fit for your lab. TEST KITS Microplate Absorbance Readers Supported: Bio-Tek ELx 800, 808 BioWhittaker Kinetic-QCL LabSystems, Multiscan, Thermo (most models) Molecular Devices EMax, VMax, ThermoMax 5 Microsoft and Excel are either registered trademarks or trademarks of Microsoft Corporation in the United States and/or other countries.

6 TEST KITS 6 Diagnostic Test Kits Test Kit Cat. No. Configuration Tests Assay Time Anaplasma Antibody Test stripwell plates 182 Kit, celisa stripwell plates minutes Babesia caballi Antibody stripwell plates 182 Test Kit, celisa stripwell plates minutes Babesia equi Antibody Test stripwell plates 182 Kit, celisa stripwell plates minutes Bluetongue Virus Antibody stripwell plates 184 Test Kit, celisa solid plates minutes Bluetongue Virus Antibody Test Kit, AGID AGID minutes* Bovine Leukemia Virus stripwell plate 91 Antibody Test Kit, ELISA stripwell plates minutes Bovine Spongiform Encephalopathy Antigen Test Kit, IHC Brucella Antibody Test Kit, ELISA Caprine Arthritis- Encephalitis Virus Antibody Test Kit, celisa Equine Infectious Anemia Virus Antibody Test Kit, ELISA Equine Infectious Anemia Virus Antibody Test Kit, AGID Escherichia coli Antigen Test Kit, K99 Pilitest TM Malignant Catarrhal Fever Virus, celisa Neospora caninum Antibody Test Kit, celisa Quick Reference Guide 298 Immunohistochemistry 50 4 hours stripwell plates stripwell plates stripwell plates stripwell plates stripwell plate stripwell plates minutes 110 minutes 35 minutes AGID minutes* 10 Latex Agglutination minutes stripwell plate 91 3 hours stripwell plates stripwell plates minutes *Incubation period is 24 hours. Sensitivity and Specificity in Perspective Relative sensitivity and specifi city values are calculated from data generated by diagnostic laboratory fi eld testing. These values are provided as guidelines only and should not be construed as the absolute sensitivity and specifi city of the test in question for any population subset.

7 Anaplasma Antibody Test Kit, celisa TEST KITS Cat. No. Species Sample Sensitivity Specificity Assay Time Configuration Tests stripwell plates 182 cattle serum 95% 98% 130 minutes stripwell plates 455 Setting a New Standard in the Diagnosis of Anaplasmosis VMRD s Anaplasma Antibody Test Kit is a competitive enzyme-linked immunosorbent assay (celisa) for the detection of antibodies specific for Anaplasma in bovine serum samples. It is intended to provide results which will give guidance about the presence of Anaplasma infection in bovine species. Sensitivity and specificity are more than four-fold better than the complement fixation test (CFT) which was the former gold standard test. In the study presented in the sensitivity and specificity table on this page, CFT was able to detect only ~20% of positive samples in three independent laboratories. Our Anaplasma celisa is a breakthrough in diagnosis of anaplasmosis in persistently-infected animals and is recommended by the OIE. It will detect antibodies to Anaplasma marginale, Anaplasma ovis, and Anaplasma centrale. The kit is available in 2-plate and 5-plate formats; both formats use breakaway stripwells. 1 Place 70 μl of samples and controls into wells of Adsorption Plate 2 Incubate 30 minutes at room temperature 3 Transfer 50 μl of samples and controls into wells of Antigen Plate 4 Incubate 60 minutes at room temperature 5 Wash two times with Wash Solution 6 Add 50 μl of Conjugate 7 Incubate 20 minutes at room temperature 8 Wash four times with Wash Solution 9 Add 50 μl of Substrate Solution 10 Incubate 20 minutes at room temperature 11 Add 50 μl of Stop Solution 12 Read at nm Samples producing <30% inhibition are negative. Samples producing 30% inhibition are positive. Formula for calculating % inhibition: % I = [(Sample OD x 100) (Mean Negative Control OD)] USDA Licensed Overview of the Anaplasma Kit Procedure For the test to be valid, the mean OD of the Negative Control must range from 0.40 to The percent inhibitioin of the Positive Control must be 30%. 7 Nested PCR + Sum VMRD Anaplasma celisa Sum Sensitivity: 95% Specifi city: 98% In-house data submitted to USDA in support of licensure, February About Anaplasmosis Anaplasmosis is a non-contagious, arthropod-borne parasitic disease of ruminants that results in significant economic losses to the cattle industry. The disease in cattle is caused by Anaplasma marginale, recently classified in geno-group II of Ehrlichiae. Anaplasma marginale is an intra-erythrocytic parasite that causes severe anemia, abortion, weight loss, jaundice and death. Diagnosis of the acute disease is based upon clinical signs, anemia and finding of Anaplasma inclusion bodies in erythrocytes. Animals surviving the acute phase become lifelong carriers. Ticks transmit the infection from carriers to naive cattle, which develop clinical disease. Cycles of rickettsemia in carriers fluctuate between and 10 7 infected erythrocytes per ml, levels generally undetectable by Giemsa staining. Carriers can be identified by detection of serum antibodies to A. marginale. See Sensitivity and Specificity In Perspective on page 6. KIT CONTENTS Component A Antigen-Coated Plates 2 plates 5 plates B Coated Adsorption/Transfer Plates 2 plates 5 plates C Positive Control 3.6 ml 3.6 ml D Negative Control 3.6 ml 3.6 ml E 100X Antibody-Peroxidase Conjugate 0.3 ml 0.5 ml F Conjugate Diluting Buffer 30 ml 60 ml G 10X Wash Solution Concentrate 120 ml 2 x 120 ml H Substrate Solution 30 ml 60 ml I Stop Solution 30 ml 60 ml An insert with Setup Record for recording sample identifi cations and results.

8 Babesia caballi Antibody Test Kit, celisa Cat. No. Species Sample Sensitivity Specificity Assay Time Configuration Tests stripwell plates 182 equine serum see below see below 105 minutes stripwell plates 455 USDA Licensed TEST KITS VMRD s Babesia caballi Antibody Test Kit, celisa and VMRD s Babesia equi Antibody Test Kit, celisa are competitive enzyme-linked immunosorbent assays which detect antibodies in equine sera to B. caballi or B. equi respectively. Antibody to B. caballi or B. equi in sample serum inhibits binding of primary monoclonal antibody. The binding of primary monoclonal antibody to the antigencoated plate is detected by binding of horeseradish peroxidase (HRP)-labeled secondary antibody. Finally, binding of the HRP-labeled secondary antibody is quantified by the addition of enzyme substrate and subsequent color product development. Strong color development indicates little or no inhibition of primary monoclonal antibody binding and therefore the absence of B. caballi or B. equi antibody in sample sera. Weak color development due to inhibition of the primary monoclonal antibody binding to the antigen on the antigen-coated plate indicates the presence of B. caballi or B. equi antibodies in sample sera. Sensitivity and Specificity of VMRD Equine Piroplasmosis Kits Based on the work of Knowles, 1 Kappmeyer, 2 and Katz 3 celisas have recently been adopted by OIE as prescribed tests for equine piroplasmosis. Two protocols developed at NVSL, one for B. caballi and one for B. equi, were validated for OIE using a 36-sample panel provided to cooperating international equine piroplasmosis reference laboratories. VMRD s piroplasmosis celisa kits are derived from these protocols and, when tested against the NVSL protocol with the same validation panel, gave 100% correct results (see Tables 1 and 3). cont. on page 9 Overview of the B. caballi Kit Procedure 1 Place 50 μl of diluted samples and controls into wells of plate 2 Incubate 30 minutes at room temperature 3 Wash three times with Wash Solution 4 Add 50 μl of Primary Antibody 5 Incubate 30 minutes at room temperature 6 Wash three times with Wash Solution 7 Add 50 μl of Secondary Antibody-HRP Conjugate 8 Incubate 30 minutes at room temperature 9 Wash three times with Wash Solution 10 Add 50 μl of Substrate Solution 11 Incubate 15 minutes at room temperature 12 Add 50 μl of Stop Solution 13 Read at nm Samples producing 40% inhibition are positive. Samples producing <40% inhibition are negative. Formula for calculating % inhibition: % I = [(Sample OD x 100) (Mean Negative Control OD)] For the test to be valid, the mean of the Negative Controls must produce an OD >0.300 and < The mean of the Positive Controls must produce an inhibition 40%. Table 1. Babesia caballi OIE check set VMRD celisa NVSL celisa + Sum Sum Sensitivity: 100% Specifi city: 100% Table 2. Babesia caballi import testing samples VMRD celisa NVSL celisa + Sum Sum Sensitivity: 100% Specifi city: 100% See Sensitivity and Specificity In Perspective on page 6. 1 Knowles, D.P., et al. Antibody to a recombinant merozoite protein epitope identifies horses infected with Babesia equi. J. Clin. Microbiol. (30): (1992). 2 Kappmeyer, L.S., et al. Detection of equine antibodies to Babesia caballi recombinant B. caballi rhoptry-associated protein 1 in a competitive-inhibition enzyme-linked immunosorbent assay. J. Clin. Microbiol. (37): (1999). 3 Katz J., et al. Procedurally similar competitive immunoassay systems for the serodiagnosis of Babesia equi, Babesia caballi, Trypanosoma equiperdum and Burkholderia mallei infection in horses. J. Vet. Diagn. Invest. (12):46 50 (2000).

9 Babesia equi Antibody Test Kit, celisa Cat. No. Species Sample Sensitivity Specificity Assay Time Configuration Tests stripwell plates 182 equine serum see below see below 105 minutes stripwell plates 455 USDA Licensed TEST KITS cont. from page 8 In the spring and summer of 2005, NVSL conducted sideby-side tests comparing the VMRD kits with the NVSL protocols. Tables 2 and 4 show the results of that testing. In late August of 2005, NVSL adopted the VMRD kits as its primary screening tests for equine piroplasmosis import testing. Overview of the B. equi Kit Procedure 1 Place 50 μl of diluted samples and controls into wells of plate 2 Incubate 30 minutes at room temperature 3 Wash three times with Wash Solution 4 Add 50 μl of Primary Antibody 5 Incubate 30 minutes at room temperature 6 Wash three times with Wash Solution 7 Add 50 μl of Secondary Antibody-HRP Conjugate 8 Incubate 30 minutes at room temperature 9 Wash three times with Wash Solution 10 Add 50 μl of Substrate Solution 11 Incubate 15 minutes at room temperature 12 Add 50 μl of Stop Solution 13 Read at nm Samples producing 40% inhibition are positive. Samples producing <40% inhibition are negative. Formula for calculating % inhibition: % I = [(Sample OD x 100) (Mean Negative Control OD)] For the test to be valid, the mean of the Negative Controls must produce an OD >0.300 and < The mean of the Positive Controls must produce an inhibition 40%. Table 3. Babesia equi OIE check set VMRD celisa NVSL celisa + Sum Sum Sensitivity: 100% Specificity: 100% Note: CFT was positive on only 3 of 12 samples positive by both VMRD and NVSL celisas. Table 4. Babesia equi import testing samples VMRD celisa NVSL celisa + Sum ** 21 1* Sum Sensitivity: 95% Specificity: 99.5% KIT CONTENTS Component A Antigen-Coated Plates 2 plates 5 plates B Positive Control 2 ml 4 ml C Negative Control 2 ml 4 ml D 100X Primary Antibody 300 μl 500 μl E 100X Secondary Antibody-Peroxidase 300 μl 500 μl F Antibody Diluting Buffer 60 ml 120 ml G Serum Diluting Buffer 9 ml 25 ml H 10X Wash Solution Concentrate 120 ml 2 x 120 I Substrate Solution 30 ml 60 ml J Stop Solution 30 ml 60 ml An insert with Setup Record for recording sample identifi cations and results. Note: CFT was positive on only 4 of 19 samples positive by both VMRD and NVSL celisas; CFT was positive on 1 sample negative by both VMRD and NVSL celisas. *39.4% inhibition by the VMRD celisa (0.6% below positive). **One sample 64.5% inhibition by the NVSL celisa (6.4% below positive); 1 sample 70.8% inhibition by NVSL celisa (0.1% below positive). 9 See Sensitivity and Specificity In Perspective on page 6.

10 Bluetongue Virus Antibody Test Kit, celisa Cat. No. Species Sample Sensitivity Specificity Assay Time Configuration Tests stripwell plates 184 ruminants serum 100% 99% 40 minutes solid plates 460 USDA Licensed TEST KITS VMRD s competitive enzyme-linked immunosorbent assay (celisa) detects antibody to bluetongue virus in ruminant sera. It has been demonstrated to detect all 24 known serotypes of Bluetongue Virus (BTV) and not to detect antibody to serotypes 1 or 2 of Epizootic Hemorrhagic Disease Virus (EHDV). The kit has demonstrated excellent sensitivity and specificity in comparison with various benchmarks in several studies. The economics of this competitively-priced assay are further enhanced by savings in technician time since sample dilution is unnecessary and incubation times total only 40 minutes. Another economic advantage of this test kit is its USDA-approved 18-month shelf life also a testimony to the stability of the kit. VMRD has manufactured over 52,000 BTV celisa plates over five million test wells. About Bluetongue Virus Bluetongue is an infectious, non-contagious, arthropodborne viral disease of wild and domestic ruminants. In cattle it is usually a subclinical infection, while in sheep it is often characterized by acute catarrhal inflammation of mucous membranes and hyperemia of coronary bands. Degenerative changes are present in skeletal and coronary musculature, which lead to weakness, prolonged convalescence and significant economic losses. Bluetongue Virus (BTV) belongs to the genus Orbivirus, family Reoviridae. Laboratory diagnosis of bluetongue is primarily established by isolation of the virus. Virus is isolated in Veros or BHK 21 cells, and its presence is confirmed by immunofluorescence. Serological methods used in diagnosis of this disease are AGID, ELISA, celisa and immunofluorescence. Positive results confirm exposure to BTV but not necessarily carrier status. Overview of the Bluetongue Kit Procedure 1 Place 25 μl of samples and controls into wells of Antigen Plate 2 Incubate 15 minutes at room temperature 3 Add 25 μl of of Conjugate 4 Incubate 15 minutes 5 Wash three times with Wash Solution 6 Add 50 μl of Substrate Solution 7 Incubate 10 minutes at room temperature 8 Add 50 μl of Stop Solution 9 Read at nm Samples are positive if they produce an OD less than 50% of the mean of the Negative Controls. Samples are negative if they produce an OD greater than or equal to 50% of the mean of the Negative Controls. For test validation, the mean OD of the Negative Controls must be greater than and less than The mean OD of the Positive Controls must be less than or equal to 50% of the mean OD of the Negative Controls. Bluetongue Virus Antibody Test Kit, AGID Cat. No. Species Sample Sensitivity Specificity Assay Time Configuration Tests ruminants serum 100% 99% 30 minutes* AGID 100 USDA Licensed Overview of Procedure VMRD s Bluetongue Virus agar gel immunodiffusion (AGID) test detects precipitating antibodies to bluetongue virus in sera of ruminants. Antibodies to Epizootic Hemorrhagic Disease Virus (EHDV) are also detected. If positive, test sera will form a line that fuses with reference lines or that cause deviation of the positive reference lines inward near the test serum well without necessarily forming a visible line. Negative sera will neither form a line nor cause deviation of the positive reference lines. 10 See Sensitivity and Specificity In Perspective on page 6. *Incubation period is 24 hours.

11 TEST KITS Bovine Leukemia Virus Antibody Test Kit, ELISA Cat. No. Species Sample Sensitivity Specificity Assay Time Configuration Tests stripwell plate 91 cattle serum 98% 100% 60 minutes stripwell plates 455 VMRD has developed a highly-sensitive and specific enzyme-linked immunosorbent assay (ELISA) kit which detects antibodies to Bovine Leukemia Virus (BLV) glycoprotein 51 (gp51) in bovine sera. Sample serum antibodies bind to BLV gp51 molecules attached to the plastic wells of the microtiter plate. Binding of these serum antibodies is detected by reaction with horseradish peroxidase (HRP)-labeled affinity-purified goat antibodies to bovine immunoglobulins. Attached HRP-labeled antibodies are detected by addition of enzyme substrate and quantitated by subsequent blue color product development. Strong color development indicates the presence of antibody to BLV gp51 in the sample serum. Very weak or no color development indicates the absence of antibody to BLV gp51 in the sample serum. VMRD s Bovine Leukemia Virus Antibody Test Kit is USDA-approved for export testing and is available in breakaway stripwell format. The assay requires that an ELISA reader be used for accurate results. KIT CONTENTS Component A Antigen-Coated Plates 1 plate 5 plates B Positive Control 3.6 ml 3.6 ml C Negative Control 3.6 ml 3.6 ml D 100X Antibody-Peroxidase Conjugate 150 μl 500 μl E Conjugate Diluting Buffer 14 ml 60 ml F 10X Wash Solution Concentrate 120 ml 2 x 120 ml G Serum Diluting Buffer 30 ml 2 x 120 ml H Substrate Solution 20 ml 60 ml I Stop Solution 20 ml 60 ml An insert with Setup Record for recording sample identifi cations and results. About Bovine Leukosis Enzootic Bovine Leukosis (EBL) is an infectious, noncontagious viral disease of cattle. It is caused by Bovine Leukemia Virus (BLV), an oncogenic delta retrovirus, which results in proliferation of B lymphocytes. Infection with BLV may lead to persistent lymphocytosis and in some adult cattle to the development of tumors with associated symptoms. The spread of disease from the introduction into a herd may take enzootic proportions. Transmission of BLV occurs between animals primarily by transfer of B lymphocytes. Overview of the BLV Kit Procedure 1 Dilute serum samples 1:25 with Serum Diluting Buffer 2 Place 50 μl of each sample into wells of the Antigen Plate 3 Incubate 20 minutes at room temperature 4 Wash all wells three times with Wash Solution 5 Add 50 μl of Conjugate 6 Incubate 20 minutes at room temperature 7 Wash three times with Wash Solution 8 Add 50 μl of Substrate Solution 9 Incubate 20 minutes at room temperature 10 Add 50 μl of Stop Solution 11 Read at nm All samples with mean OD values greater than or equal to the mean OD of the Positive Controls are positive for BLV. All samples with mean OD values less than the mean of the Positive Controls are negative for BLV. For test validation, the mean OD of the Negative Controls must be less than The mean OD of the Positive Controls must be and < Reference ELISA + Sum USDA Licensed VMRD BLV 4* ELISA Sum Sensitivity: 98% Specifi city: 100% * All calves less than 8 months of age. Based on a specifi c sample set. However, no diagnostic test kit is always 100% specifi c on all sample populations. Since market introduction of our BLV kit, occasional false positives have been encountered. We therefore advise all users that when BLV prevalence is low, positive samples should be confi rmed by some other method, particularly where valuable animals may be involved and/or when BLV status is used as the single criterion for disposition of animals. Whenever import restrictions do not prohibit it, VMRD will provide reference assay service for positives of high-value animals or for positives in low-prevalence situations. We are not capable of testing large numbers of samples, and therefore cannot provide this reference assay service for all positives found. Data generated by three independent laboratories during fi eld trial testing of VMRD s BLV ELISA as required for USDA licensure, February Trauma, use of common bleeding needles and surgical procedures are the main means of transmission. It is rarely vertically transmitted. Most BLV infections are inapparent. Approximately 5% of animals develop clinical signs. AGID and ELISA tests are used to identify carrier cattle. Control programs for EBL include testing and removal of positive animals. Several European countries which have instituted eradication programs also require that imported cattle be free of BLV. See Sensitivity and Specificity In Perspective on page 6.

12 Brucella Antibody Test Kit, ELISA Cat. No. Species Sample Sensitivity Specificity Assay Time Configuration Tests stripwell plates 184 multiple serum see below see below 70 minutes stripwell plates 460 TEST KITS Works With Cattle, Goat and Sheep Sera! VMRD s Brucella Antibody Test Kit is an indirect enzymelinked immunosorbent assay (ELISA) that detects antibodies to Brucella abortus and B. melitensis in bovine and small ruminant sera. This kit s high sensitivity and specificity make it an excellent tool for eradication programs. Background is very low, and most positive samples give very strong signals. Assay time is only 70 minutes. The conjugate detects bovine, caprine, and ovine IgG, enabling the kit to be used for all three species. caprine samples Defi ned Samples + Sum VMRD Brucella ELISA Sum Sensitivity: 100% Specificity: 99.8% Overview of the Brucella Kit Procedure 1 Dilute serum samples 1:100; milk is tested undiluted 2 Add 100 μl of samples and controls into Antigen Plate 3 Incubate 30 minutes at room temperature 4 Wash four times with Wash Solution 5 Add 100 μl of Conjugate 6 Incubate 30 minutes at room temperature 7 Wash four times with Wash Solution 8 Add 100 μl of Substrate Solution 9 Incubate 10 minutes at room temperature 10 Add 100 μl of Stop Solution 11 Read at nm Samples having <25% SP ratio are negative. Samples having 25% SP ratio are positive. Formula for calculating SP ratio: SP = [(Sample OD - Negative Control OD) (Mean Positive Control OD - Mean Negative Control OD)] x 100 The mean OD of the Negative Controls must be < The OD of the Positive Controls must be and bovine samples Defi ned Samples + Sum VMRD Brucella ELISA Sum Sensitivity: 100% Specificity: 99.3% ovine samples Defi ned Samples + Sum VMRD Brucella ELISA Sum Sensitivity: 100% Specifi city: 97.3% About Brucellosis Brucellosis is an infectious disease caused by bacteria of the genus Brucella. Brucellosis is commonly transmitted to susceptible animals by direct contact with infected animals or with an environment that has been contaminated with discharges from infected animals. Aborted fetuses, placental membranes or fluids, and other vaginal discharges present after an infected animal has aborted or calved are all highly contaminated with infectious Brucella organisms. The disease may also be spread when wild animals or animals from an affected herd mingle with brucellosisfree herds. There is no effective way to detect infected animals by their appearance. The most obvious signs in pregnant animals are abortion or birth of weak calves. Brucellosis is diagnosed in the laboratory by isolating Brucella organisms in various samples or by detecting antibodies against the bacteria in serum or milk samples. Considering the damage done by the infection decreased milk production, weight loss in animals, loss of young, infertility, and lameness brucellosis is one of the most serious diseases of livestock. The rapidity with which it spreads and the fact that it is transmissible to humans makes it all the more serious.* Not Available For Sale In the US 12 See Sensitivity and Specificity In Perspective on page 6. * Portions of this paragraph were taken from the APHIS website at:

13 TEST KITS Caprine Arthritis-Encephalitis Virus Antibody Test Kit, celisa Cat. No. Species Sample Sensitivity Specificity Assay Time Configuration Tests stripwell plates 184 caprine serum 100% 99.6% 110 minutes stripwell plates 460 The study of CAEV has a long history at VMRD. Dr. Scott Adams, President of VMRD, was a member of the team that initially isolated CAEV and characterized the disease in the late 1970s and early 1980s. VMRD s competitive enzyme-linked immunosorbent assay (celisa) detects antibodies to caprine arthritisencephalitis virus (CAEV) in goat sera. Our CAEV celisa test utilizes a proprietary xeno-monoclonal antibody derived by fusion of goat splenocytes and mouse myeloma cells which has excellent characteristics for use in celisa. This antibody is conjugated to horeseradish peroxide and is used to compete with serum antibodies for antigen bound to the microtiter plate. Most indirect ELISA systems presently in use lack specificity to varying degrees. False positive reactions are particularly undesirable in goats of high commercial value. VMRD s competitive ELISA assay for CAEV antibody detection eliminates most of these non-specific reactions. Several validation studies, in addition to the one summarized here, have confirmed the superior quality of VMRD s CAEV celisa test kit. Overview of the CAEV Kit Procedure 1 Place 50 μl of samples and controls into wells of Antigen Plate 2 Incubate 60 minutes at room temperature 3 Wash three times with Wash Solution 4 Add 50 μl of Conjugate 5 Incubate 30 minutes at room temperature 6 Wash three times with Wash Solution 7 Add 50 μl of Substrate Solution 8 Incubate 20 minutes at room temperature 9 Add 50 μl of Stop Solution 10 Read at nm Samples producing <35% inhibition are negative. Samples producing 35% inhibition are positive. Formula for calculating % inhibition: % I = [(Sample OD x 100) (Mean Negative OD)] USDA Licensed For the test to be valid, the mean OD of the Negative Controls must be The mean of the Positive Controls must produce 35% inhibition. About Caprine Arthritis-Encephalitis Caprine Arthritis-Encephalitis (CAE) is one of the most important diseases of goats worldwide. Two manifestations of disease occur: Encephalitis in young kids 2 to 4 months of age, and arthritis in adult goats. CAE is caused by a lentivirus closely related to North American isolates of Ovine Progressive Pneumonia Virus (OPPV). The infection persists for life and antibodies to Caprine Arthritis Encephalitis Virus (CAEV) can be detected using, among others, AGID, ELISA and IFA procedures. The major mode of transmission is from doe to kid through milk and colostrum. KIT CONTENTS Component A Antigen-Coated Plates 2 plates 5 plates B Positive Control 3.6 ml 3.6 ml C Negative Control 3.6 ml 3.6 ml D 100X Antibody-Peroxidase Conjugate 300 μl 500 μl E Conjugate Diluting Buffer 30 ml 60 ml F 10X Wash Solution Concentrate 120 ml 2 x 120 ml G Substrate Solution 30 ml 60 ml H Stop Solution 30 ml 60 ml An insert with Setup Record for recording sample identifi cations and results. caprine samples CAEV AGID and IP + Sum VMRD CAEV celisa Sum Sensitivity: 100% Specifi city: 99.6% Field Testing Data, See Sensitivity and Specificity In Perspective on page 6. Herrmann, L.M., et al. Competitive-inhibition enzyme-lin ked immunosorbent assay for detection of serum antibodies to caprine arthritis-encephalitis virus: Diagnostic tool for successful eradication. Clin. Diagn. Lab. Immunol. 10(2): (2003). [This publication documents that the celisa was more sensitive than immunoprecipitation for detecting antibody in sera of experimentally-infected goats.]

14 Equine Infectious Anemia Virus Antibody Test Kit, AGID TEST KITS Cat. No. Species Sample Sensitivity Specificity Assay Time Format Tests equine serum 99% 100% 30 minutes AGID 200 VMRD s Equine Infectious Anemia Virus (EIAV) agar gel immunodiffusion (AGID) test detects precipitating antibodies in sera of Equidae to purified recombinant EIAV core protein of 26,000 molecular weight (p26). Using highly purified recombinant p26 protein antigen reduces problems of interpretation associated with extraneous precipitin lines from contamination by non-relevant antigens. The antigenantibody precipitation reaction takes place in agar gel using the 7-well standard procedure developed by John W. Black and described by Pearson (American Association of Veterinary Laboratory Diagnosticians, 22nd Annual Proceedings, pp , 1979). Purified soluble EIAV p26 antigen is placed in the center well and reference positive control serum is placed in three alternating peripheral wells. Sample sera are placed in the three remaining wells. After incubation, reference lines form between the antigen well and the reference positive control serum wells. Sample sera, if positive, will form a line that fuses with reference positive control lines or that deviate the reference positive control lines inward near the sample well without formation of a visible line. Negative sera will neither form a line that fuses with the reference positive control line nor cause deviation of the reference positive control lines. Always in Stock. No Back Orders. About Equine Infectious Anemia Equine Infectious Anemia (EIA) is caused by a lentivirus. It produces acute episodes of disease that are interspersed with clinically normal periods. The acute episodes usually last for a few days and are associated with fever, thrombocytopenia, and anemia. In most infected horses, the disease episodes occur with less and less frequency until an inapparent carrier state develops. The infection is life long and, if stressed, inapparent carrier horses may express recurrent viremia and disease. Transmission occurs by transfer of blood from one horse to another by biting insects or contaminated needles and instruments. Transmission is most likely during episodes of clinical disease when the virus titer is highest in the blood, and is least likely during the carrier stage. Unfortunately, it is difficult to know at what stage an infected horse may be and when another episode might occur. It can be diagnosed by detection of antibody to the capsid p26 protein of the virus. This internal viral protein is relatively conserved among EIA virus strains, allowing detection of antibody in virtually all infected horses. VMRD EIA AGID Reference Assay + Sum Sum Sensitivity: 99% Specifi city: 100% Composite of all Field Tests, USDA Licensed See Sensitivity and Specificity In Perspective on page 6. EIAV Testing Regulations For USA Customers: VMRD, in compliance with Federal regulations, will only ship EIAV test kits to USDAapproved laboratories. The sale and use of EIAV test kits in the USA is restricted to laboratories approved by State and Federal (USDA) animal health officials. The National Veterinary Services Laboratories will periodically supply coded check test samples to evaluate the competency of the USDA-approved laboratories. For questions about becoming an EIAV-licensed testing lab, contact the USDA. 14

15 TEST KITS Equine Infectious Anemia Virus Antibody Test Kit, ELISA Cat. No. Species Sample Sensitivity Specificity Assay Time Configuration Tests stripwell plate 94 equine serum 100% 100% 35 minutes stripwell plates 470 VMRD s Equine Infectious Anemia Virus (EIAV) enzymelinked immunosorbent assay (ELISA) detects antibodies to EIAV p26 in equine sera. EIAV-specific antibodies bind to recombinant p26 antigen when sera are added to the test wells. Unbound antibody is washed away and antigen-bound antibody is detected by the addition of horseradish-peroxidase (HRP)-conjugated p26 antigen. Unbound conjugate is washed away and, with the addition of substrate solution, wells containing antibody turn blue. When stop solution is added, positive wells turn yellow and negative wells remain clear. Test results may be calculated from optical density values reported by a microplate absorbance reader or determined visually by comparing color development intensity of sample wells with that of the positive control. VMRD s EIAV ELISA is rapid and convenient only 35 minutes total incubation time, no sample dilution, and only two washes yet it is highly specific and sensitive. In-House Testing 1628 AGID-negative samples tested negative on VMRD s ELISA. It correctly classified all samples in the 2001, 2002, 2003, 2004, and the 2005 USDA CVB EIAV check sets. On a panel of 10 weak samples, VMRD s assay correlated 100% with the consensus of three USDAlicensed EIAV ELISAs. Field Testing In field trails conducted by three independent laboratories on a total of 543 samples, VMRD s new ELISA had 100% correlation with the various reference assays used. As a test of robustness, a tester independency panel consisting of 25 samples 12 positive and 13 negative was distributed to the participating laboratories. All were in agreement on all samples with the exception of one laboratory that called one negative sample positive. Overview of the EIAV ELISA Kit Procedure 1 Place 50 μl of samples and controls into wells of Antigen Plate 2 Incubate 10 minutes at room temperature 3 Wash once with Wash Solution 4 Add 50 μl of Conjugate 5 Incubate 10 minutes at room temperature 6 Wash four times with Wash Solution 7 Add 50 μl of Substrate Solution 8 Incubate 15 minutes at room temperature 9 Add 50 μl of Stop Solution to all wells 10 Read at 450 nm or by eye Samples are positive if they produce an OD greater than or equal to the mean of the positive control. Samples are negative if they produce an OD less than the mean of the positive control. For the test to be valid, the OD of the Positive Control should be greater than or equal to 1.5 times the OD of the Negative Control. The OD of the Negative Control should be less than or equal to For the test to be valid when reading by eye, the Positive Control should have visible yellow color and the Negative Control should have no or faint visible color that is less than the Positive Control. Samples producing positive test results are to be sent in to the National Veterinary Services Laboratories (NVSL) for verification. VMRD EIA ELISA Reference Assay + Sum Sum Sensitivity: 100% Specifi city: 100% Composite of all Field Tests, USDA Licensed VMRD s ELISA sensitivity is comparable or superior to other USDA-licensed ELISAs on titrations of positive samples and in detection of weak samples. VMRD s kit contains no thimerosal and generates no hazardous waste. KIT CONTENTS Component A Antigen-Coated Plates 1 plate 5 plates B Positive Control 2 ml 4 ml C Negative Control 2 ml 4 ml D 100X Antibody-Peroxidase Conjugate 150 μl 500 μl E Conjugate Diluting Buffer 15 ml 60 ml F 10X Wash Solution Concentrate 60 ml 2 x 120 ml G Substrate Solution 15 ml 60 ml H Stop Solution 15 ml 60 ml An insert with Setup Record for recording sample identifi cations and results. 15 See Sensitivity and Specificity In Perspective on page 6.

16 Escherichia coli Antigen Test Kit (K99 Pilitest ) Cat. No. Species Sample Sensitivity Specificity Assay Time Format Tests cattle serum 93% (<48 hr) 75% 10 minutes Latex Agglutination USDA Licensed TEST KITS VMRD S K99 PILITEST Ten percent of beef and dairy calves are lost each year to scours. One of the most important infectious causes of scours in young calves is enteropathogenic Escherichia coli which carries K99 + pili. Fortunately, immunoprophylaxis has been shown to be effective in reducing the severity of scours caused by enteropathogenic E. coli. Nevertheless, because of the sporadic nature of outbreaks, the expense of vaccines and oral antibodies may not be justified without a specific etiologic diagnosis of K99 + Escherichia coli. Our K99 Pilitest is a sensitive latex agglutination test for the presence of K99 + E. coli in stools of diarrheic calves. It may also be used for detection of K99 + E. coli grown on solid or in liquid medium. The agglutination reaction takes less than three minutes and the entire test, including sample preparation, can be performed in ten minutes or less. VMRD s Pilitest is recommended for use with stools from calves exhibiting signs of diarrhea or white scours within 5 days of birth. While it is possible to detect K99 + E. coli in stools of older calves, its contribution to diarrhea in this group may not be significant. Bacterial numbers fall in some calves after 48 hours. Therefore, when one is testing calves having diarrhea for more than 48 hours, it may be necessary to test several calves before concluding that K99 + E. coli are not responsible for the diarrhea observed. Overview of the K99 Kit Procedure 1 Prepare sample in accordance with insert 2 Shake all four squeeze bottles. 3 Add one drop of solution A to first three rings on the black 4 Add one drop of solution D to the fourth ring of the black slide 5 Add a drop of the sample to the first and the fourth rings 6 Add a drop of solution B to the second ring 7 Add a drop of solution C to the third ring 8 Mix the two drops in each ring with separate stir sticks 9 Gently rock the slide for two minutes observe for agglutination 10 Allow slide to remain still for one minute 11 Observe for agglutination Agglutination in ring one indicates infection with K99 + E. coli as long as no agglutination occurs in rings three and four. If no agglutination occurs in ring one within three minutes it is considered negative provided agglutination does occur in ring two. Wash the black slide with water and wipe dry between tests. <48 hours post-inoculation VMRD K99 PILITEST TM Culture + Sum * 14 1 ** 3 4 Sum Sensitivity: 93% Specificity: 75% * Sample was from an inoculated calf. Three consecutive cultures following the negative culture were positive. Given that the data support this sample being positive, diagnostic specifi city of this assay on this sample set was 100%. ** A previous sample from the same animal (taken at <24 hours) tested positive by the Pilitest. TM 16 See Sensitivity and Specificity In Perspective on page 6. KIT CONTENTS Component A Antibody-Coated Latex Beads 1.75 ml 4.0 ml B Positive Control 0.75 ml 1.9 ml C Negative Control 0.75 ml 1.9 ml D Latex Bead Control 0.75 ml 1.9 ml E Black Slide with 4 rings and stir sticks 1 1 Analytical Sensitivity and Specificity: In an independent evaluation, VMRD s K99 Pilitest TM detected the following standard E. coli K99 isolates at a concentration of 3.6 x 10 9 CFU per ml: B41, B44, B79, B85, B111, and B117. The test also detected the following E. coli K99 field isolates at a concentration of 3.6 x 10 9 CFU per ml: 833, 1678, 678, 8425B, 635B, and Isolates , B41M, 1592, , , and were used as negative controls in this study. All were negative in the Pilitest TM with the exception of isolate 1592, which was weakly positive at a concentration of 8.2 x CFU per ml (1000 times the detection level of K99 bacteria).

17 Malignant Catarrhal Fever Virus, celisa Cat. No. Species Sample Sensitivity Specificity Assay Time Configuration Tests 294 multiple serum see below see below 3 hours 1 stripwell plate 91 TEST KITS VMRD s competitive enzyme-linked immunosorbent assay (celisa) detects malignant catarrhal fever (MCF) antibodies in ruminant sera or plasma. It is a specific, rapid, monoclonal-antibody-based assay for detection of antibody against the MCF group viruses. It was developed using a monoclonal antibody to an epitope conserved among all MCF group viruses examined to date. These viruses include alcelaphine herpesvirus 1, alcelaphine herpesvirus 2, ovine herpesvirus 2, caprine herpesvirus 2, the virus of unknown origin causing classic MCF in white-tailed deer, and the MCF viruses carried by musk ox, ibex and oryx. VMRD s celisa test kit utilizes a monoclonal antibody (15-A-AC) derived by fusion of mouse myeloma cells and the splenocytes from the mouse hyperimmunized with MCF viral antigens (Minnesota isolate). The antibody is conjugated to horseradish peroxide and used to compete with antibodies in the test serum for antigen previously bound to the microtiter plate. Overview of the MCFV Kit Procedure 1 Dilute samples and controls 1:5 with Diluting Buffer 2 Add 50 μl of samples and controls into Antigen Plate 3 Incubate 60 minutes at room temperature 4 Wash three times with Wash Solution 5 Add 50 μl of Conjugate 6 Incubate 60 minutes at room temperature 7 Wash three times with Wash Solution 8 Add 100 μl of Substrate Solution 9 Incubate 60 minutes at room temperature 10 Add 100 μl of Stop Solution 11 Read at 450 nm Samples causing <25% inhibition are negative. Samples causing 25% inhibition are positive. Formula for calculating % inhibition: % I = [(Sample OD x 100) (Mean Negative Control OD)] The mean OD of the Negative Control must range from 0.40 to The inhibition of the Positive Control must be 25%. Sensitivity About Malignant Catarrhal Fever Malignant catarrhal fever (MCF) is a severe, frequently fatal disease syndrome primarily of ruminant species. It is caused by one of a group of Rhadinoviruses in the Gammaherpesvirus subfamily. These viruses exist in nature as well-adapted, inapparent infections in certain ruminants that act as reservoir hosts. Virtually all members of a carrier species are infected under natural conditions. Based on the reservoir ruminant species from which the causative virus arises, several epidemiologic forms of MCF are recognized: Wildebeest-associated MCF, sheep-associated MCF, and the recently reported goat-associated MCF. The disease is transmitted only between carriers and clinically-susceptible species. Significant percentages of clinically-susceptible hosts can be subclinically infected. celisa + celisa Total % Cattle with clinical MCF Bison with clinical MCF Sheep Specificity celisa + celisa Total % Cattle, bison, deer Sheep Bison Not Available for Sale in the US 17 Hong Li et al. Unpublished data, April See Sensitivity and Specificity In Perspective on page 6.

18 Neospora caninum Antibody Test Kit, celisa Cat. No. Species Sample Sensitivity Specificity Assay Time Format Tests stripwell plates 184 multiple serum 96% 99% 100 minutes stripwell plates 460 USDA Licensed TEST KITS VMRD s Neospora test is a competitive enzymelinked immunosorbent assay (celisa) that detects antibodies against Neospora caninum in cattle sera. Our competitive ELISA format allows other species to be tested, but validation has been completed only on cattle. An immunodominant surface protein of 65 kda is captured on the antigen plate using a monoclonal antibody. Another horseradish peroxidase-conjugated monoclonal anti body competes with serum antibodies for a specific epitope on p65. Sensitivity and specificity studies on both species confirm the high accuracy of this kit. In a mass screening of 4323 sera of unknown serologic status only 5% of sera fell within ±5% of the cut-off value, confirming a clear distinction between positive and negative sera bimodal distribution. VMRD s Neospora kit is available in a 2-plate and 5-plate format with breakaway stripwells Overview of the Neospora Kit Procedure 1 Add 50 µl of samples and controls into Antigen Plate 2 Incubate 60 minutes at room temperature 3 Wash three times with Wash Solution 4 Add 50 µl of Conjugate 5 Incubate 20 minutes at room temperature 6 Wash three times with Wash Solution 7 Add 50 µl of Substrate Solution 8 Incubate 20 minutes at room temperature 9 Add 50 µl of Stop Solution 10 Read at nm Samples producing <30% inhibition are negative. Samples producing 30% inhibition are positive. Formula for calculating % inhibition: % I =100 - [(Sample OD x 100) (Mean Negative Control OD)] For the test to be valid, the mean OD of the Negative Control must be 0.30 and <2.50. The inhibition of the Positive Control must be 30%. Bovine samples Reference Assay + Sum VMRD Neospora celisa Sum Sensitivity: 96% Specifi city: 99% 18 KIT CONTENTS Component A Antigen-Coated Plates 2 plates 5 plates B Positive Control 3.6 ml 3.6 ml C Negative Control 3.6 ml 3.6 ml D 100X Antibody-Peroxidase Conjugate 300 μl 500 μl E Conjugate Diluting Buffer 30 ml 60 ml F 10X Wash Solution Concentrate 120 ml 2 x 120 ml G Substrate Solution 30 ml 60 ml H Stop Solution 30 ml 60 ml An insert with Setup Record for recording sample identifi cations and results. See Sensitivity and Specificity In Perspective on page 6. VMRD celisa Field Testing, About Neosporosis Neosporosis is a recently-described disease that has been identified across the world in various species, including dogs, cattle, sheep, goats, and horses. It is caused by Neospora caninum, a protozoan parasite closely related to Toxoplasma gondii. Even though the dog can be the definitive host for Neospora, it is not known if there are other definitive hosts. No signs of clinical illness are noted in cows that abort due to Neospora either prior to the abortion or post-abortion. The aborted fetuses are usually autolyzed with no gross lesions and placentas are not retained. Abortions have been diagnosed in both heifers and cows from 3 months gestation to term. A majority (78%) of Neospora abortions occur between 4 and 6 months gestation. This pattern of mid-gestation abortion is distinct from other diagnosed causes of infectious abortion in dairy cattle which tend to occur later in gestation. In dogs, Neospora infection causes neuromuscular paralysis. Identification of carrier animals is based upon detection of specific antibody with serological tests while diagnosis of abortions is based upon microscopic examination of the fetus and immunohistochemistry.

19 Bovine Spongiform Encephalopathy Antigen Test Kit, Immunohistochemistry USDA Licensed for Export Cat. No. Species Sample Sensitivity Specificity Assay Time Tests 298 bovine obex 100% 100% 4 hours 50 TEST KITS VMRD s Bovine Spongiform Encephalopathy (BSE) Antigen Test Kit provides a standard operating procedure for detection of prion protein (PrP) in brain and lymphoid tissues of bovines using monoclonal antibody immunohistochemistry. Antibody F99/ recognizes a conserved epitope (QYQRES) of the ruminant prion protein. VMRD s BSE test kit contains all critical reagents necessary to perform the assay. It includes target retrieval solution, antibody diluent, antibody F99/97.6.1, anti-mouse biotinylated secondary antibody, peroxidaselabeled streptavidin and AEC substrate. Not Available for Sale in the US About Transmissible Spongiform Encephalopathies Transmissible Spongiform Encephalopathies (TSEs) are fatal neurodegenerative diseases. Included among them are Bovine Spongiform Encephalopathy (BSE) of cattle, Scrapie of sheep and goats, and Chronic Wasting Disease (CWD) of mule deer and elk. They are caused by prion proteins (proteinaceous infectious particles) that lack nucleic acid. Prions are composed largely, if not entirely, of an abnormal isoform of a normal cellular protein. TSEs occur worldwide. Laboratory diagnoses of TSEs are made by histopathology, ELISA, Western blot, and immunohistochemistry (IHC). The unique advantage of the latter is its ability to confirm specificity by architectural histologic distribution of prions. No other procedure currently available can do this. 19 See Sensitivity and Specificity In Perspective on page 6.

20 FA REAGENTS 20

21 Immunofluorescence Reagents Fluorescent antibody (FA) techniques, direct and indirect, are standby procedures that remain unsurpassed for versatility and accurate detection of either antigen or antibodies. The FA technique offers rapid deployment of new assays with a minimum of development time. It has the distinct advantage over other assay methods of enabling the operator to visually distinguish between specific and non-specific reactions. Essential equipment and facilities to perform FA: Quality epifluorescence microscope with a mercury or xenon lamp. The microscope must be located in a dark room to obtain optimum visualization. Other necessary equipment standard for biomedical laboratories. We provide positive and negative controls for as many of our IFA systems as possible. We try to make all of our positive controls at an antibody concentration two to four two-fold dilutions above endpoint to avoid an excessively strong positive control contaminating a critical sample. We take great pains with every step of conjugate production --from antibody development to purification to conjugation-- to make sure that our FITC conjugates are indeed pathogen-specific. We try to produce consistent products; however, immunofluorescence reagents still tend to vary from lot to lot. Therefore, we provide a detailed certificate of analysis unique to each lot informing you what to expect. This certificate also provides useful information such as the strain of the pathogen, screening dilution, and recommended procedure for performing the assay. A picture is worth a thousand words, so we include a lotspecific photograph of a positive reaction on as many of our certificates of analysis as possible. We are always ready and willing to work through any problem that you may encounter. We also consult on all aspects of fluorescent antibody assays from setting up an FA lab to designing a custom FA system. In addition, we are also willing to look at your prepared slides and to ship slides that we have prepared for your perusal. We can also referee difficult samples and send pictures with comments to aid with future tough calls. Feel free to call or us whenever you have an FA question. FA Reagents VMRD s immunofluorescence reagents are set apart by quality, consistency and standardization. After more than fifteen years of using and selling immunofluorescence reagents, we strive to use that experience and knowledge to make your life easier. We optimize the dilution of our secondary antibody conjugates for use in all of the applicable IFA systems that we sell. All of our anti-pathogen conjugates and positive and negative controls are ready to use so that you don t waste time and reagents determining the optimal dilution. We test our diluents in all of our systems in which they might be used to avoid problems with background, nonspecificity or lack of signal. VMRD Facility, Pullman, Washington USA

22 FA REAGENTS VMRD CARRIES THE MOST EXTENSIVE RANGE OF VETERINARY FLUORESCENT ANTIBODY PRODUCTS AVAILABLE ANYWHERE INDIRECT IMMUNOFLUORESCENCE Indirect immunofluorescence is used to detect antibodies in body fluids of diseased animals. Materials for indirect immunofluorescence (IFA) include: FA Substrate Slide Contains 10 or 12 wells spotted with an antigen. Positive & Negative Controls Used on each slide for the purpose of comparison. Serum Diluting Buffer Used to dilute samples to working dilution. (Catalog No SB or SB) Anti-Immunoglobulin Conjugate Used to detect bound antibody on the slide. (see p. 29) Rinse Buffer Used for washing off unbound antibodies and conjugates. (Catalog No RB) Mounting Fluid Used to enhance visualization of fluorescence. (Catalog No MF) Recommended procedure for indirect immunofluorescence (IFA): 1. Warm slide to room temperature before removing from foil pouch. 2. Dilute serum in serum diluting buffer, ph 7.2. Place µl* of diluted serum on the designated wells. 3. Incubate slide in humid chamber at 37 C for 30 minutes. 4. Using a wash bottle, gently rinse slide briefly in FA rinse buffer, ph 9.0, and then soak for 10 minutes in FA rinse buffer, ph Drain slide and dry around wells by pressing blotter (included in pouch) to front surface. Place µl* FITC-labeled anti-igg or -IgM conjugate on the wells. 6. Incubate as in step Rinse as in step Drain slide and dry back and edges with a paper towel. Do not allow stained surface to dry. Do not rinse with water. 9. Mount with mounting fluid and view with a good quality fluorescence microscope at X. Confirmation may be made at 400X. DIRECT IMMUNOFLUORESCENCE Direct immunofluorescence is used to detect antigens. Materials for direct immunofluorescence (FA) include: Direct FA Conjugate Antibodies conjugated to FITC. Control Slide Used to check performance of a conjugate. Contains two wells: one positive and one negative. Rinse Buffer Used for washing off unbound antibodies and conjugates. (Catalog No RB) Mounting Fluid Used to enhance visualization of fluorescence. (Catalog No MF) Recommended procedure for direct immunofluorescence (FA): 1. Warm slide to room temperature before removing from foil pouch. 2. Place 50 µl of direct FA conjugate on the designated wells. 3. Incubate slide in humid chamber at 37 C for 30 minutes. 4. Using a wash bottle, gently rinse slide briefly in FA rinse buffer, ph 9.0, and then soak for 10 minutes in FA rinse buffer, ph Drain slide and dry back and edges with a paper towel. Do not allow stained surface to dry. Do not rinse with water. 6. Mount with mounting fluid and view with good quality fluorescence microscope at X. 7. Confirmation may be made at 400X. Clostridium chauvoei Clostridium novyi Clostridium septicum Clostridium sordellii This 12-well FA substrate slide contains three patterns of four wells. Each pattern contains a well spotted with Clostridium chauvoei, a well spotted with Clostridium novyi, a well spotted with Clostridium septicum and a well spotted with Clostridium sordellii bacteria = C. chauvoei. 2 = C. novyi. 3 = C. septicum. 4 = C. sordellii * Volumes depend on the size of the wells; see product insert for specific volumes.

23 Bovine Immunofluorescence Reagents Infectious Agent Product Type (Origin) Size Catalog No. Positive Blood Slide 12 well BBI Babesia bigemina Positive Control for IFA (bovine) 1 ml 211-P-BBI Negative Control for IFA (bovine) 1 ml 211-N-BBI Positive Blood Slide 12 well BBO Babesia bovis Positive Control for IFA (bovine) 1 ml 211-P-BB Negative Control for IFA (bovine) 1 ml 211-N-BB FA Substrate Slide 10 well BT FA REAGENTS Bluetongue Virus (BTV) Bovine Adenovirus Type 3 (BAV3) Bovine Adenovirus Type 5 (BAV5) Bovine Coronavirus (BCV) Positive Control for IFA (bovine) 1 ml 211-P-BTV Negative Control for IFA (bovine) 1 ml 211-N-BTV FITC Conjugate (murine) 1 ml BT FITC Conjugate (murine) 10 ml BT FITC Conjugate (caprine) 1 ml BAV3 FITC Conjugate (caprine) 10 ml BAV3 FITC Conjugate (bovine) 1 ml BAV5 FITC Conjugate (bovine) 10 ml BAV5 FITC Conjugate (bovine) 1 ml BCV FITC Conjugate (bovine) 10 ml BCV Bovine Herpesvirus Type 1/ Infectious Bovine Rhinotracheitis (BHV1/IBR) Bovine Leukemia Virus (BLV) Bovine Parainfluenza Virus Type 3 (PI3) Bovine Parvovirus (BPV) FA Control Slide 2 well IBR FA Substrate Slide 10 well IBR Positive Control for IFA (bovine) 1 ml 211-P-IBR Negative Control for IFA (bovine) 1 ml 211-N-IBR FITC Conjugate (caprine) 1 ml IBR FITC Conjugate (caprine) 10 ml IBR FA Control Slide 2 well BLV FA Substrate Slide 10 well BLV Positive Control for IFA (bovine) 1 ml 211-P-BLV Negative Control for IFA (bovine) 1 ml 211-N-BLV FA Control Slide 2 well PI3 FA Substrate Slide 10 well PI3 Positive Control for IFA (bovine) 1 ml 211-P-PI3 Negative Control for IFA (bovine) 1 ml 211-N-PI3 FITC Conjugate (caprine) 1 ml PI3 FITC Conjugate (caprine) 10 ml PI3 FITC Conjugate (caprine) 1 ml BPV FITC Conjugate (caprine) 10 ml BPV 23

24 Bovine Immunofluorescence Reagents Infectious Agent Product Type (Origin) Size Catalog No. FA Control Slide 2 well BRSV Bovine Respiratory Syncytial Virus (BRSV) FA Substrate Slide 10 well BRSV Positive Control for IFA (bovine) 1 ml 211-P-BRSV Negative Control for IFA (bovine) 1 ml 211-N-BRSV FITC Conjugate (caprine) 1 ml BRSV FITC Conjugate (caprine) 10 ml BRSV FA Control Slide 2 well BVD FA Substrate Slide 10 well BVD Bovine Viral Diarrhea Virus (BVDV) Positive Control for IFA (bovine) 1 ml 211-P-BVD Negative Control for IFA (bovine) 1 ml 211-N-BVD FITC Conjugate (porcine) 1 ml BVD FA REAGENTS FITC Conjugate (porcine) 10 ml BVD Clostridium chauvoei Clostridium novyi Clostridium septicum see page 28 Clostridium sordellii Clostridium spp. 4-way Neospora caninum see page 28 Reovirus FITC Conjugate (caprine) 1 ml REO FITC Conjugate (caprine) 10 ml REO Canine Immunofluorescence Reagents Infectious Agent Product Type (Origin) Size Catalog No. Borrelia burgdorferi Lyme Disease Brucella canis Canine Brucellosis see page 28 FA Substrate Slide 12 well CB Positive Control for IFA (canine) 1 ml 211-P-CB Negative Control for IFA (canine) 1 ml 211-N-CB FA Control Slide 2 well CAV FA Substrate Slide 10 well CAV Canine Adenovirus (CAV) Positive Control for IFA (canine) 1 ml 211-P-CAV Negative Control for IFA (canine) 1 ml 211-N-CAV FITC Conjugate (porcine) 1 ml CAV FITC Conjugate (porcine) 10 ml CAV 24

25 Canine Immunofluorescence Reagents Infectious Agent Product Type (Origin) Size Catalog No. FA Control Slide 2 well CCV Canine Coronavirus (CCV) FA Substrate Slide 10 well CCV FITC Conjugate (feline) 1 ml CCV FITC Conjugate (feline) 10 ml CCV FA Control Slide 2 well CDV FA Substrate Slide 10 well CDV IgG Positive Control for IFA (canine) 1 ml 211-P-CDV-G IgM Positive Control for IFA (canine) 1 ml 211-P-CDV-M Canine Distemper Virus (CDV) Polyclonal FITC Conjugate (caprine) 1 ml CDV Polyclonal FITC Conjugate (caprine) 10 ml CDV Monoclonal FITC Conjugate (murine) 1 ml CDV FA REAGENTS Canine Herpesvirus (CHV) Monoclonal FITC Conjugate (murine) 10 ml CDV Positive Blood Smear each 214-P-CDV Negative Blood Smear each 214-N-CDV FA Substrate Slide 10 well CHV Positive Control for IFA (canine) 1 ml 211-P-CHV FITC Conjugate (canine) 1 ml CHV FITC Conjugate (canine) 10 ml CHV FA Control Slide 2 well CPI Canine Parainfluenza Type 2 (CPI2) Canine Parvovirus (CPV) Ehrlichia canis Leishmania infantum FA Substrate Slide 10 well CPI Positive Control for IFA (canine) 1 ml 211-P-CPI Negative Control for IFA (canine) 1 ml 211-N-CPI FITC Conjugate (porcine) 1 ml CPI2 FITC Conjugate (porcine) 10 ml CPI2 FA Control Slide 2 well CPV FA Substrate Slide 10 well CPV IgG Positive Control for IFA (canine) 1 ml 211-P-CPV-G IgM Positive Control for IFA (canine) 1 ml 211-P-CPV-M FITC Conjugate (murine) 1 ml CPV FITC Conjugate (murine) 10 ml CPV FA Substrate Slide 12 well EC Positive Control for IFA (canine) 1 ml 211-P-EC Negative Control for IFA (canine) 1 ml 211-N-EC FA Substrate Slide 12 well LSH Positive Control for IFA (canine) 1 ml 211-P-LSH Negative Control for IFA (canine) 1 ml 211-N-LSH Neospora caninum see page 28 25

26 Canine Immunofluorescence Reagents Infectious Agent Product Type (Origin) Size Catalog No. Rabies Recombinant Nucleoprotein Rickettsia rickettsii Rocky Mountain Spotted Fever (RMSF) see page 28 FA Substrate Slide 12 well RMSF Positive Control for IFA (canine) 1 ml 211-P-RMSF Negative Control for IFA (canine) 1 ml 211-N-RMSF Equine Immunofluorescence Reagents Infectious Agent Product Type (Origin) Size Catalog No. FA REAGENTS Anaplasma phagocytophila (formerly Ehrlichia equi) Equine Herpesvirus Type 1/ Equine Rhinopneumonitis Virus (EHV1/ERV) FA Substrate Slide 12 well EE Positive Control for IFA (equine) 1 ml 211-P-EE Negative Control for IFA (equine) 1 ml 211-N-EE FA Control Slide 2 well ERV FA Substrate Slide 10 well ERV Positive Control for IFA (equine) 1 ml 211-P-ERV-EQ Positive Control for IFA (llama) 1 ml 211-P-ERV-LL FITC Conjugate (caprine) 1 ml ERV FITC Conjugate (caprine) 10 ml ERV Neorickettsia risticii FA Substrate Slide 12 well NR Rabies Recombinant Nucleoprotein see page 28 Feline Immunofluorescence Reagents Infectious Agent Product Type (Origin) Size Catalog No. Bartonella henselae Chlamydia psittaci Feline Chlamydiosis FA Substrate Slide 12 well BH IgG Positive Control for IFA (feline) 1 ml 211-P-BH-G IgM Positive Control for IFA (feline) 1 ml 211-P-BH-M FA Substrate Slide 12 well FC FA Control Slide 2 well FCV Feline Calicivirus (FCV) FA Substrate Slide 10 well FCV Positive Control for IFA (feline) 1 ml 211-P-FCV Negative Control for IFA (feline) 1 ml 211-N-FCV FA Control Slide 2 well FVR Feline Herpesvirus/ Feline Viral Rhinotracheitis (FHV/FVR) FA Substrate Slide 10 well FVR Positive Control for IFA (feline) 1 ml 211-P-FHV FITC Conjugate (feline) 1 ml FVR 26 FITC Conjugate (feline) 10 ml FVR

27 Feline Immunofluorescence Reagents Infectious Agent Product Type (Origin) Size Catalog No. FIP FA Control Slide 2 well FIP FIP-1 FA Control Slide 2 well FIP1 FIP-2 FA Control Slide 2 well FIP2 FIP-1 FA Substrate Slide 10 well FIP1 Feline Coronaviruses Feline Infectious Peritonitis (FIP-1 & FIP-2) FIP-2 FA Substrate Slide 10 well FIP2 FIP-1 Positive Control for IFA (feline) 1 ml 211-P-FIP1 FIP-2 Positive Control for IFA (feline) 1 ml 211-P-FIP2 FIP-1 Negative Control for IFA (feline) 1 ml 211-N-FIP1 FIP-2 Negative Control for IFA (feline) 1 ml 211-N-FIP2 FITC Conjugate (feline) 1 ml FIP FITC Conjugate (feline) 10 ml FIP FA REAGENTS Feline Leukemia Virus (FeLV) Feline Panleukopenia Virus (FPLV) FA Control Slide 2 well FELV Primary Antibody for IFA (caprine) 10 ml FELV1 Secondary Antibody for IFA (equine) 10 ml FELV2 FeLV Reagent Set for IFA 2 x 10 ml FELV Positive Blood Smear each 214-P-FELV Negative Blood Smear each 214-N-FELV FA Control Slide 2 well FPL FA Substrate Slide 10 well FPL FITC Conjugate (murine) 1 ml FPL FITC Conjugate (murine) 10 ml FPL Toxoplasma gondii see page 29 Porcine Immunofluorescence Reagents Infectious Agent Product Type (Origin) Size Catalog No. Porcine Adenovirus (PAV) FITC Conjugate (porcine) 1 ml PAV FITC Conjugate (porcine) 10 ml PAV FA Control Slide 2 well PCRV FA Substrate Slide 10 well PCRV Porcine Circovirus (PCV) Positive Control for IFA (porcine) 1 ml 211-P-PCRV Negative Control for IFA (porcine) 1 ml 211-N-PCRV FITC Conjugate (porcine) 1 ml PCRV FITC Conjugate (porcine) 10 ml PCRV FA Control Slide 2 well PPV Porcine Parvovirus (PPV) FA Substrate Slide 10 well PPV FITC Conjugate (porcine) 1 ml PPV 27 FITC Conjugate (porcine) 10 ml PPV

28 Porcine Immunofluorescence Reagents Infectious Agent Product Type (Origin) Size Catalog No. FA Control Slide 2 well TGE Transmissible Gastroenteritis Virus (TGEV) FA Substrate Slide 10 well TGE Positive Control for IFA (porcine) 1 ml 211-P-TGE FITC Conjugate (feline) 10 ml 211-N-TGE FITC Conjugate (feline) 1 ml TGE FITC Conjugate (feline) 10 ml TGE Vesicular Stomatitis Virus (VSV) FITC Conjugate (porcine) 1 ml VSV FITC Conjugate (porcine) 10 ml VSV Multiple Species Immunofluorescence Reagents FA REAGENTS Infectious Agent Product Type (Origin) Size Catalog No. Borrelia burgdorferi Lyme Disease Clostridium chauvoei Clostridium novyi FA Substrate Slide 12 well LD Positive Control for IFA (canine) 1 ml 211-P-LD Negative Control for IFA (canine) 1 ml 211-N-LD FA Substrate Slide 12 well CC FITC Conjugate (caprine) 1 ml CC FITC Conjugate (caprine) 10 ml CC FA Substrate Slide 12 well CN FITC Conjugate (caprine) 1 ml CN FITC Conjugate (caprine) 10 ml CN FA Substrate Slide 12 well CS Clostridium septicum FITC Conjugate (bovine) 1 ml CS FITC Conjugate (bovine) 10 ml CS FA Substrate Slide 12 well CSO Clostridium sordellii FITC Conjugate (caprine) 1 ml CSO FITC Conjugate (caprine) 10 ml CSO Clostridium spp. 4-way FA Substrate Slide 12 well C4 FA Substrate Slide 12 well NC Positive Control for IFA (bovine) 1 ml 211-P-NC-BOV Positive Control for IFA (canine) 1 ml 211-P-NC-CAN Neospora caninum Negative Control for IFA (bovine) 1 ml 211-N-NC-BOV Negative Control for IFA (canine) 1 ml 211-N-NC-CAN FITC Conjugate (caprine) 1 ml NC FITC Conjugate (caprine) 10 ml NC Rabies Recombinant Nucleoprotein FA Control Slide 2 well RAB FITC Conjugate (equine) 1 ml RAB FITC Conjugate (equine) 10 ml RAB 28

29 Multiple Species Immunofluorescence Reagents Infectious Agent Product Type (Origin) Size Catalog No. FA Substrate Slide 12 well TOXO Toxoplasma gondii IgG Positive Control for IFA (feline) 1 ml 211-P-TOXO-FEL IgM Positive Control for IFA (feline) 1 ml 211-P-TOXO-FEL-M Negative Control for IFA (feline) 1 ml 211-N-TOXO-FEL Immunofluorescence Reagent Secondary Conjugates (Indirect) FITC Anti-Immunoglobulin Conjugates Origin Size Catalog No. caprine 1 ml Anti-Bovine IgG 1,2, affinity purified, heavy and light chains caprine 10 ml Anti-Canine IgG caprine 1 ml caprine 10 ml FA REAGENTS Anti-Canine IgG, affinity purified Anti-Canine IgM, affinity purified, heavy chain specific Anti-Equine IgG Anti-Equine IgG, affinity purified rabbit 1 ml 035-AP-1 rabbit 10 ml 035-AP-10 caprine 1 ml caprine 10 ml caprine 1 ml caprine 10 ml caprine 1 ml 043-AP-1 caprine 10 ml 043-AP-10 Anti-Feline IgG Anti-Feline IgM, affinity purified, heavy chain specific Anti-Goat IgG Anti-Llama IgG, affinity purified, heavy and light chains Anti-Mouse IgG, affinity purified Anti-Pig IgG, affinity purified, heavy and light chains ovine 1 ml ovine 10 ml caprine 1 ml caprine 10 ml rabbit 1 ml rabbit 10 ml caprine 1 ml caprine 10 ml rabbit 1 ml rabbit 10 ml rabbit 1 ml caprine 10 ml Immunofluorescence Buffers & Mounting Fluid Rinse Buffers & Mounting Fluid Size Catalog No. FA Conjugate Diluting Buffer 100 ml CB FA Serum Diluting Buffer with 1% BSA 100 ml SB FA Special Serum Diluting Buffer with 10% Bovine Serum 100 ml SB FA Mounting Fluid 10 ml MF FA Rinse Buffer, powdered (makes 4 L) 1 pkg RB

30 PUMPER, courtesy Jennifer Miller Antibodies Polyclonal Antibodies POLYCLONAL ANTISERA TO INFECTIOUS AGENTS p. 31 POLYCLONAL ANTISERA TO IMMUNOGLOBULINS p Monoclonal Antibodies MONOCLONAL ANTIBODIES TO INFECTIOUS AGENTS p MONOCLONAL ANTIBODIES TO CELL MARKERS p MONOCLONAL ANTIBODIES TO IMMUNOGLOBULINS p. 39 MONOCLONAL ANTIBODIES TO MHC COMPLEX p. 39 MONOCLONAL ANTIBODIES TO CYTOKINES AND HORMONES p. 39 ANTIBODIES Custom Antibody Production In addition to offering the antibodies listed on the following pages, VMRD, Inc. provides expert custom antibody production services to scientific investigators and clinical diagnosticians. You may have purified an antigen or have access to an antigen, yet find antiserum production inconvenient, logistically impossible, or prohibitively expensive. Our objective is to produce the highest-quality antiserum possible against the antigen you provide. Antisera are produced in a USDAapproved licensed facility and in full compliance with the Animal Welfare Act. Most of VMRD s monoclonal antibodies are produced in mice and sold as clarified and filtered ascites fluid which is preserved with sodium azide. Mouse ascites monoclonals are packaged in liquid form, usually at a concentration of 1.0 mg/ml. VMRD s monoclonals are available in three sizes: 0.1 ml; 0.5 ml; and 1.0 ml. To order simply choose the cell line and amount you require. 30

31 POLYCLONAL ANTIBODIES Polyclonal Antisera to Infectious Agents Specificity Origin Size Catalog No. Bovine Adenovirus Type 3 (BAV-3) caprine 2 ml BAV3 Bovine Herpesvirus Type 1/Infectious Bovine Rhinotracheitis Virus (BHV-1/IBR) caprine 2 ml IBR Bovine Leukemia Virus (BLV) bovine 2 ml BLV Bovine Parainfluenza Virus Type 3 (PI-3) caprine 2 ml PI3 Bovine Respiratory Syncytial Virus (BRSV) caprine 2 ml BRSV Bovine Viral Diarrhea Virus (BVDV) caprine 2 ml BVD Canine Coronavirus (CCV) feline 2 ml CCV Canine Distemper Virus (CDV) caprine 1 ml CDV Canine Parainfluenza Virus (CPI) porcine 2 ml CPI Equine Herpesvirus Type 1/Equine Rhinopneumonitis Virus (EHV-1/ERV) caprine 1 ml ERV Feline Infectious Peritonitis Virus Type 1 (FIP-1) feline 1 ml FIP1 Feline Infectious Peritonitis Virus Type 2 (FIP-2) feline 2 ml FIP2 Feline Panleukopenia Virus (FPL) canine 2 ml FPL Neospora caninum caprine 1 ml NC Porcine Circovirus (PCV) porcine 2 ml PCRV Porcine Parvovirus (PPV) porcine 1 ml PPV Toxoplasma gondii caprine 1 ml TOXO Transmissible Gastroenteritis Virus (TGEV) feline 2 ml TGE ANTIBODIES Polyclonal Antisera to Immunoglobulins Specificity Origin Size Catalog No. Anti-Bovine IgA Polyclonal Antiserum caprine 2 ml 5 ml Anti-Bovine IgG (H + L) Polyclonal Antiserum caprine 2 ml 5 ml Anti-Bovine IgG1 Polyclonal Antiserum caprine 2 ml 5 ml Anti-Bovine IgG1,2 Polyclonal Antiserum caprine 2 ml 5 ml Anti-Bovine IgG2 Polyclonal Antiserum caprine 2 ml 5 ml Anti-Bovine IgM (mu chain) Polyclonal Antiserum caprine 2 ml 5 ml Anti-Bovine Whole Polyclonal Antiserum caprine 2 ml 5 ml Anti-Canine IgA Polyclonal Antiserum caprine 2 ml 5 ml Anti-Canine IgG (H + L) Polyclonal Antiserum caprine 2 ml 5 ml Anti-Canine IgM (mu chain) Polyclonal Antiserum caprine 2 ml 5 ml Anti-Canine Whole Polyclonal Antiserum caprine 2 ml 5 ml Anti-Equine IgA Polyclonal Antiserum caprine 2 ml 5 ml

32 Polyclonal Antisera to Immunoglobulins Specificity Origin Size Catalog No. Anti-Equine IgG(a,b,c) Polyclonal Antiserum caprine 2 ml 5 ml Anti-Equine IgG (H + L) Polyclonal Antiserum caprine 2 ml 5 ml Anti-Equine IgG(T) Polyclonal Antiserum caprine 2 ml 5 ml Anti-Equine IgM (mu chain) Polyclonal Antiserum caprine 2 ml 5 ml Anti-Equine Whole Polyclonal Antiserum caprine 2 ml 5 ml Anti-Feline IgG (H + L) Polyclonal Antiserum ovine 2 ml 5 ml MONOCLONAL ANTIBODIES Monoclonal Antibodies to Infectious Agents ANTIBODIES Specificity Origin Isotype Cell Line Anaplasma marginale (MSP1) Mouse Ascites IgG3 15D2 Anaplasma marginale (MSP2) Mouse Ascites IgG1 O50A2 Babesia bigemina (p36, p20 and p16) Mouse Ascites IgG Babesia bigemina (p58) Mouse Ascites IgG Babesia bigemina (p72) Mouse Ascites IgG Bovine Herpesvirus Type 1 (BHV-1/IBR) (gb - gi) Mouse Ascites IgG2b D9E7 Bovine Herpesvirus Type 1 (BHV-1/IBRIBR) (gb - gi) Mouse Ascites IgG2b H2 Bovine Herpesvirus Type 1 (BHV-1/IBR) (gc - giii) Mouse Ascites IgG1 G2 Bovine Herpesvirus Type 1 (BHV-1/IBR) (gc - giii) Mouse Ascites IgG2b F2 Bovine Herpesvirus Type 1 (BHV-1/IBR) (gd - giv) Mouse Ascites IgG1 1B8-F11 Bovine Herpesvirus Type 5 (BHV5) (gc) Mouse Ascites IgM L6G Bovine Leukemia Virus (BLV) (gp51 - G) Mouse Ascites IgG1 BLV1 Bovine Leukemia Virus (BLV) (gp51 - D-D ) Mouse Ascites IgG1 BLV2 Bovine Leukemia Virus (BLV) (p24) Mouse Ascites IgG1 BLV3 Bovine Parainfluenza Virus Type 3 (PI3) (p69) Mouse Ascites IgG2a 1B6 Bovine Parainfluenza Virus Type 3 (PI3) (p69) Mouse Ascites IgG2a 2A2 Bovine Viral Diarrhea Virus (BVDV) (gp55) Mouse Ascites IgG2a D89 Bovine Viral Diarrhea Virus Type 1 (BVDV) E2 (gp53) Mouse Ascites IgG2a 157 Bovine Viral Diarrhea Virus Type 2 (BVDV) E2 (gp53) Mouse Ascites IgG2a BA-2 Bovine Viral Diarrhea Virus Type 2 (BVDV) E2 (gp53) Mouse Ascites IgG2a BA-29 Bovine Viral Diarrhea Virus Types 1 & 2 (BVDV) E2 (gp53) Mouse Ascites IgG1 BA-26(a) Bovine Viral Diarrhea Virus Types 1 & 2 (BVDV) E2 (gp53) Mouse Ascites IgG2b

33 Monoclonal Antibodies to Infectious Agents ANTIBODIES Specificity Origin Isotype Cell Line Canine Adenovirus Type 1 (CAV-1) Mouse Ascites IgG1 2E10-H2 Canine Adenovirus Type 2 (CAV-2) Mouse Ascites IgG2a 4H1-A7 Canine Distemper Virus (CDV) (nucleoprotein) Cell Culture IgG2b CDV-NP Canine Distemper Virus (CDV) (envelope) Mouse Ascites IgG1 1C42H11 Canine Parainfluenza Virus Type 2 (CPI-2) Cell Culture IgG2b, κ CPI-A-CA light chain Canine Parvovirus (CPV) Mouse Ascites IgG2a A3B10 Caprine Arthritis Encephalitis Virus (CAEV) (p28) (CAEV-63, CAEV-Co, MVV, OPPV) Mouse Ascites IgG1 CAEP5A1 (CAEV-63, CAEV-Co, MVV) Mouse Ascites IgG1 CAEP10A1 (CAEV-63, CAEV-Co, MVV) Mouse Ascites IgG1 CAEP8B1 (CAEV-63, CAEV-Co) Mouse Ascites IgG1 CAEP13B1 (CAEV-63) Mouse Ascites IgG1 CAEP12A1 Cryptosporidium parvum Mouse Ascites IgM Equine Arteritis Virus (EAV) (nucleocapsid) Mouse Ascites IgG1 17D3 Equine Infectious Anemia Virus (EIAV) (p26) Mouse Ascites IgG1 EIAP6A1 Leptospira interrogans serovar australis Mouse Ascites IgG3 F90 C6 Leptospira interrogans serovar canicola Mouse Ascites IgM F152 C11 Leptospira interrogans serovar copenhageni Mouse Ascites IgG1 F70 C24 Leptospira interrogans serovar hardjo Mouse Ascites IgG1 F22 C6 Leptospira interrogans serovar icterohaemorrhagiae Mouse Ascites IgG3 F70 C14 Leptospira interrogans serovar pomona Mouse Ascites IgM F48 C6 Malignant Catarrhal Fever Virus (MCFV) Mouse Ascites IgG2b 15-A-AC Malignant Catarrhal Fever Virus (MCFV) (p17) Mouse Ascites IgG2a N10-A Malignant Catarrhal Fever Virus (MCFV) (p24) Mouse Ascites IgG1 36-A Malignant Catarrhal Fever Virus (MCFV) (p36/34) Mouse Ascites IgM N55-A Neospora caninum (gp65) Mouse Ascites IgG1 5B6 Porcine Parvovirus (PPV) Mouse Ascites IgG1 3C9D11H11 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) (nucleocapsid) Mouse Ascites IgG2b 2D6 Prion Protein (IHFG) Mouse Ascites IgG1 F89/ Prion Protein (QYQRES) Mouse Ascites IgG1 F99/ Pseudorabies Virus (gp50) Mouse Ascites IgG2b 6D8-MB4 Psuedorabies Virus (giii) Mouse Ascites IgG2b 3G9F3 6D8-MB4 A3B10 CDV-NP D89 D89 Pseudorabies Virus (gp50) Canine Parvovirus Canine Distemper Virus (nucleoprotein) Bovine Viral Diarrhea Virus (gp55) on Type 1 Bovine Viral Diarrhea Virus (gp55) on Type 2 33

34 Monoclonal Antibodies to Cell Markers ANTIBODIES Specificity Cell Line Isotype Cattle Goat Sheep Swine Horse Cat Dog Ferret Mink Human Mouse Rabbit Prothymocytes EqT12 IgM nt nt nt nt nt nt nt Thymocytes EqT7 IgG1 nt nt nt nt + nt nt nt nt nt nt nt Thymocytes EqT6 IgG3 nt nt nt nt + nt nt nt nt nt nt nt Thymocyte subpopulation 25-2E IgM nt nt nt nt nt + nt nt nt nt nt nt Thymocyte subpopulation 25-2F IgM nt nt nt nt nt + nt nt nt nt nt nt Thymocyte subpopulation 25-2G IgM nt nt nt nt nt + nt nt nt nt nt nt Thymocytes + (αβ + γδ) T subpopulations 2B11 IgM nt nt nt + nt nt nt nt nt nt nt nt T Lymphocytes (Pan T1) CF54A UgG nt - - nt T Lymphocytes (Pan T1) RTH2A IgG1 nt nt nt nt nt nt nt nt nt nt nt + T Lymphocytes (Pan T1) RTH230A IgG1 nt nt nt nt nt nt nt nt nt nt nt + T Lymphocytes (Pan T2) CF74A IgG1 nt nt nt nt nt + nt nt nt nt nt nt T Lymphocytes (Pan T2) CF255A IgG nt nt - - nt T Lymphocytes (Pan T2) RTH21A IgG1 nt nt nt nt nt nt nt nt nt nt nt + T Lymphocytes (Pan T2) RT22A IgM nt nt nt nt nt nt nt nt nt nt nt + T Lymphocytes + Basophils (Pan T3) MRB61A IgG1 nt nt nt nt nt nt nt nt nt nt nt + T Lymphocytes (Pan T4) RTH26A IgG2a nt nt nt nt nt nt nt nt nt nt nt + T Lymphocytes (Pan T4) RTH65A IgM nt nt nt nt nt nt nt nt nt nt nt + Pan T Lymphocytes 25-2I IgG1 nt nt nt nt nt + nt nt nt nt nt nt Pan T Lymphocytes 25-1A IgM nt nt nt nt nt nt nt nt nt + nt nt Pan Lymphocytes PG89A IgG1 nt nt nt + nt nt nt nt nt nt nt nt Pan Lymphocytes RTH186A IgG1 nt nt nt nt nt nt nt nt nt nt nt + Pan Lymphocytes MRB102A IgM nt nt nt nt nt nt nt nt nt nt nt + Pan Lymphocytes PG106A IgM nt nt nt + nt nt nt nt nt nt nt nt Lymphocyte subpopulation DH52A IgM nt nt nt nt nt nt + nt nt nt nt nt T Lymphocyte subpopulation CF7A IgG * + nt nt nt T Lymphocyte subpopulation, Granulocytes ISC24A IgM nt nt nt nt nt nt nt nt nt nt nt + T Lymphocyte subpopulation, Granulocytes (CD45R0-like) ISC4A IgG3 nt nt nt nt nt nt nt nt nt nt nt + T Lymphocytes (γδ), CD4 + CD8 subpopulations PG74C1 IgM nt nt nt + nt nt nt nt nt nt nt nt T + B Lymphocytes E47B IgM nt nt WC7-N1 (T + B Lymphocytes) TH18A IgG nt nt T + B Lymphocytes, Monocytes CF26A IgG nt nt nt nt T + B Lymphocyte subpopulations CAT54A IgM nt nt nt nt nt + nt nt nt nt nt nt T + B Lymphocyte subpopulations, Granulocytes E38A IgG1 nt nt nt nt nt nt nt T + B Lymphocyte subpopulations, Granulocytes HB72A IgG2a nt nt nt nt nt nt nt Basophils, CD4 + T Lymphocyte subpopulation RACT20A IgG1 nt nt nt nt nt nt nt nt nt nt nt + Basophils, Granulocytes, Monocytes MRB120A IgG1 nt nt nt nt nt nt nt nt nt nt nt + B Lymphocytes, Monocytes, Granulocytes and Platelets F72A IgG nt - nt nt CD IgG2a - nt nt + nt nt nt nt nt nt nt nt CD1b TH97A IgG2a nt nt nt nt CD2 BAQ95A IgG nt nt CD2 CH128A IgG nt nt CD2 8-1E3 IgG1 + nt nt nt nt nt nt nt nt nt nt nt CD2 HB88A IgG nt nt nt CD2 16-1E10 IgG2a + nt nt nt nt nt nt nt nt nt nt nt CD2 36F-18 IgG2a nt nt + nt nt nt nt nt nt nt nt nt CD2 MSA4 IgG2a nt nt nt CD2 (sheep red blood cell receptor) MUC2A IgG2a nt nt CD2 PG168A IgG3 nt nt nt + nt nt nt nt CD2 B26A4 IgM nt nt CD3 MM1A IgG nt nt nt nt - nt nt CD3 8E6 IgG nt nt nt nt nt - nt nt 34

35 Monoclonal Antibodies to Cell Markers ANTIBODIES Specificity Cell Line Isotype Cattle Goat Sheep Swine Horse Cat Dog Ferret Mink Human Mouse Rabbit CD3 8C8 IgG2a nt nt nt + nt nt nt nt nt nt nt nt CD3 2B3C IgG2b nt nt nt + nt nt nt nt nt nt nt nt CD4 RTH1A IgG1 nt nt nt nt nt nt nt nt nt nt nt + CD4 CACT138A IgG nt nt CD4 17D1 IgG nt nt nt nt nt nt nt nt CD4 HB61A IgG nt - CD4 CAT30A IgG nt - CD4 GC1A IgG2a nt nt nt nt - nt nt CD4 IL-A11 IgG2a nt nt nt nt nt nt nt nt CD4 PT90A IgG2a nt - CD IgG2b nt nt - nt - CD4 GC50A1 IgM nt - CD4 CACT83B IgM nt nt CD4 DH29A IgM nt nt CD4 and CD8 subpopulations (CD28-like) RACT19A IgM nt nt nt nt nt nt nt nt nt nt nt + CD5 CACT105A IgG nt nt nt CD5 GR60A IgG nt nt nt CD5 PG114A IgG1 nt nt nt + - nt nt nt nt nt nt nt CD5 9G12 IgG1 nt nt nt + nt nt nt nt nt nt nt nt CD5 HT23A IgG nt nt CD5 DH3B IgG nt CD5 HB19A IgG2a nt nt nt nt - nt nt CD5 in cattle, B cells only in horses B29A IgG2a nt nt CD5 BAGB145A IgM nt nt nt CD5 DH13A IgM nt CD5 DH14A IgM nt nt CD6 PG90A IgG nt nt nt nt nt nt nt CD6 BAQ91A IgG1 + +* +* nt nt CD6 BAQ82A IgM + - +* nt nt CD8α CACT80C IgG nt nt CD8α PT36B IgG nt nt CD8α IgG2a nt nt nt + nt nt nt nt nt nt nt nt CD8α PT81B IgG2b nt nt CD8α 73/6.9.1 IgG nt nt nt nt nt nt nt CD8α BAQ111A IgM + +* nt nt CD8β BAT82A IgG nt nt CD8β PG164A IgG2a nt nt nt + nt nt nt nt nt nt nt nt CD8 HT14A IgG nt nt CD8 25-2A IgG1 nt nt nt nt nt + nt nt nt nt nt nt CD8 CADO46A IgG1 nt nt nt nt nt nt nt nt nt CD8 ISC29E IgG1 nt nt nt nt nt nt nt nt nt nt nt + CD8 ISC38A IgG1 nt nt nt nt nt nt nt nt nt nt nt + CD8 25-1B IgG2a nt nt nt nt nt nt nt nt nt + nt nt CD8 ISC27A IgG2a nt nt nt nt nt nt nt nt nt nt nt + 35 *polymorphic

36 ANTIBODIES Monoclonal Antibodies to Cell Markers ANTIBODIES Specificity Cell Line Isotype Cattle Goat Sheep Swine Horse Cat Dog Ferret Mink Human Mouse Rabbit CD8 ISC16A IgM nt nt nt nt nt nt nt nt nt nt nt + CD11a F IgG1 nt nt + nt nt nt nt nt nt nt nt nt CD11a MUC76A IgG2a nt nt nt nt CD11b MM12A IgG nt nt nt nt nt nt nt nt CD11b MM10A IgG2b nt nt nt nt nt nt nt nt CD11b BAQ147A IgM nt nt nt nt nt nt nt nt CD11b RT18A IgM nt nt nt nt nt nt nt nt nt nt nt + CD11c BAQ153A IgM nt nt nt CD11c RT3A IgM nt nt nt nt nt nt nt nt nt nt nt + CD13-like (Monocytes/Neutrophils) 25-2B IgG1 nt nt nt nt nt + nt nt nt nt nt nt CD14 (M-M8) MM61A IgG nt nt - nt nt CD14 (M-M9) CAM36A IgG nt + CD18 BAQ30A IgG nt + CD18 BAT75A IgG nt nt nt CD18 H20A IgG * nt nt + nt + CD21 BB6-11C9 IgG1 nt nt nt + nt nt nt nt nt nt nt nt CD21 BAQ15A IgM nt nt nt nt nt nt nt nt CD21-like (B-B7) GB25A IgG nt nt nt nt nt nt nt nt CD21-like (Cr-B1) F46A IgG nt nt nt CD24-like (Granulocytes + B Lymphocytes) MUC106A IgM nt nt nt + nt nt nt nt nt nt nt nt CD25 (Bovine Interleukin-2 receptor α) CACT108A IgG2a nt nt nt nt nt CD25 (Bovine Interleukin-2 receptor α) CACT116A IgG nt nt nt nt nt CD25 (Bovine Interleukin-2 receptor α) LCTB2A IgG nt nt nt nt nt nt nt nt CD25 (Porcine Interleukin-2 receptor) PGBL25A IgG1 nt nt nt + nt nt nt nt nt nt nt nt CD26 CACT114A IgG2b nt nt nt nt nt CD29 FW4-101 IgG nt nt nt nt nt + nt nt CD41/61 (platelets) CAPP2A IgG nt nt nt nt nt nt nt nt CD41/61 CL2A IgG nt nt nt nt nt CD44 PORC24A IgG2a nt CD44 + Endothelium BAT31A IgG nt nt CD44 + Endothelium BAG40A IgG nt + CD45-like CADO18A IgG1 nt nt nt nt nt nt + nt nt nt nt nt CD45-like CADO19A IgG1 nt nt nt nt nt nt + nt nt nt nt nt CD45 BAGB20A IgG nt nt nt CD45 ISC39A IgG1 nt nt nt nt nt nt nt nt nt nt nt + CD45 ISC18A IgG2a nt nt nt nt nt nt nt nt nt nt nt + CD45 CACTB51A IgG2a nt nt nt nt nt nt nt CD A1 IgM nt nt nt + nt nt nt nt nt nt nt nt CD45 ISC76A IgM nt nt nt nt nt nt nt nt nt nt nt + CD45R GS5A IgG nt nt nt nt nt nt nt nt CD45R LCT27A IgG nt nt nt nt nt nt nt nt CD45R LCT2A IgG2a nt nt nt nt nt nt nt nt CD45R GC6A IgM nt nt nt nt nt nt nt nt CD45RA PG96A IgG1 nt nt nt + nt nt nt nt nt nt nt nt 36

37 Monoclonal Antibodies to Cell Markers ANTIBODIES Specificity Cell Line Isotype Cattle Goat Sheep Swine Horse Cat Dog Ferret Mink Human Mouse Rabbit CD45RA PG167A IgG1 nt nt nt + nt nt nt nt nt nt nt nt CD45RA PGB78A IgG2a nt nt nt + nt nt nt nt nt nt nt nt CD45RA (p220) 73B1 IgG1 nt nt + nt nt nt nt nt nt nt nt nt CD45RB DH16A IgM nt - + nt - CD45R0 GC42A1 IgG nt nt nt nt nt nt nt nt CD45R0 GC44A IgG nt nt nt nt nt nt nt nt nt CD45R0 IL-A116 IgG3 + nt nt nt nt nt nt nt nt nt nt nt CD47 TH17A IgM nt nt nt nt nt + nt nt CD49d FW3-218 IgG2b nt nt nt nt nt nt nt nt CD58 RTH33A IgG1 nt nt nt nt nt nt nt nt nt nt nt + CD58 RTH32A IgM nt nt nt nt nt nt nt nt nt nt nt + CD62L BAQ92A IgG nt nt nt nt nt nt nt nt CD62L (L Selectin) DU1-29 IgG nt nt nt nt nt nt nt nt CD90 (Thy-1) DH2A IgM nt nt CD90 (Thy-1) (reacts only with granulocytes in horse) DH24A IgM nt nt nt CD172a (SWC3) DH59B IgG1 + +* +* nt nt CD172a (SWC3) IgG1 nt nt nt + nt nt nt nt nt nt nt nt CD172a (SWC3) A IgG2b + +* nt + nt nt - WC1 Pan Leukocytes IL-A29 IgG nt nt WC1-N1 Pan Leukocytes B7A1 IgM nt nt WC1-N2 Pan Leukocytes BAQ4A IgG nt nt nt WC1-N3 (γδ T Lymphocyte subpopulation) CACTB32A IgG1 + +* +* - nt nt nt nt WC1-N3 Pan Leukocyte cluster BAQ72A IgM nt nt nt nt WC1-N4 Pan Leukocyte cluster BAQ159A IgG nt nt WC1-N4 Pan Leukocyte cluster BAG25A IgM nt nt nt WC1-N4 Pan Leukocyte subpopulation BAQ89A IgG nt nt nt WC1-N5 Pan Lymphocyte subpopulation BAQ136A IgM + nt nt nt nt WC1-N11 Pan Leukocyte cluster BAQ167A IgG nt nt nt WC1-N11 Pan Leukocyte subpopulation BAQ90A IgG nt nt nt nt WC1-N22 Pan Leukocytes CACTB31A IgG2b nt nt nt nt WC1-N25 Pan Leukocytes GB54A IgG2a nt nt nt nt nt nt nt WC1-N26 Pan Leukocytes GB45A IgG nt nt nt nt nt nt nt nt WC15 (Red Blood Cells) ANA8A IgG1 + nt nt nt nt nt nt nt nt nt nt nt Neutrophils CADO48A IgG1 nt nt nt nt nt nt + nt nt nt nt nt TcR1-N1 subpopulation PT79A IgG2a nt nt TcR1-N4 (δ chain) PGBL22A IgG1 nt nt nt + nt nt nt nt nt nt nt nt TcR1-N6 (γδ T Lymphocyte subpopulation) CACTB6A IgM nt nt nt nt TcR1-N6 cluster (γ chain) CACTB14A IgG nt nt nt nt TcR1-N7 cluster (γ chain subpopulation) 86D IgG nt nt nt nt nt nt nt nt TcR1-N7 (γδ chain) CACTB81A IgG nt nt nt nt TcR1-N12 (δ chain) CACT61A IgM nt nt nt nt TcR1-N21 (δ chain) CACT148A IgM nt TcR1-N24 (δ chain) GB21A IgG2b + + nt nt nt nt nt nt nt nt nt nt Pan B Lymphocytes RACT30A IgM nt nt nt nt nt nt nt nt nt nt nt + 37 * polymorphic

38 Monoclonal Antibodies to Cell Markers ANTIBODIES Specificity Cell Line Isotype Cattle Goat Sheep Swine Horse Cat Dog Ferret Mink Human Mouse Rabbit Pan B Lymphocytes MRB29A IgM nt nt nt nt nt nt nt nt nt nt nt + B Lymphocytes MRB143A IgM nt nt nt nt nt nt nt nt nt nt nt + B Lymphocytes (B-B1) BAS9A IgM nt nt B Lymphocytes (B-B2) BAQ44A IgM nt + B Lymphocytes (B-B4) BAQ155A IgG nt nt B Lymphocytes (B-B5) CH127A IgM nt nt B Lymphocytes (B-B6) GC65A IgM nt nt nt B Lymphocytes (B-B11) LCT30A IgG nt nt nt nt nt nt nt nt B Lymphocytes (B-B14) LCTB16A IgG nt nt nt nt nt nt nt nt B Lymphocyte subpopulation E18A IgG2a nt nt B Lymphocyte subpopulation CADO34A IgM nt nt nt nt nt nt + nt nt nt nt nt B Lymphocyte subpopulation RT19A IgM nt nt nt nt nt nt nt nt nt nt nt + B Lymphocyte subpopulation (ileal Peyer s patch B Lymphocytes) BB6-10A10 IgM - nt nt + nt nt nt nt nt nt nt nt B + T Lymphocyte subpopulations RACT14A IgM nt nt nt nt nt nt nt nt nt nt nt + B + T Lymphocyte subpopulations RACT21A IgM nt nt nt nt nt nt nt nt nt nt nt + Monocytes (M-M1) PG130A IgM nt nt nt nt nt nt nt nt Monocytes (M-M2) CH137A IgM nt nt Monocytes (M-M3) MM29A IgM nt nt nt nt nt Monocytes (M-M7) BAQ151A IgG nt nt nt nt nt SWC5 (γδ T Lymphocyte subpopulation) PG92A IgM nt nt nt γδ T Lymphocyte subpopulation PG94A IgM nt nt nt Activation Molecule 1 (αβ + γδ T cells) CACT101A IgM nt nt nt nt nt Activation Molecule 2 (predominant reactivity with γδ T cells) CACT26A IgG nt nt nt nt nt Activation Molecule 2 (predominant reactivity with γδ T cells) CACT63A IgG nt nt nt nt nt Activation Molecule 2 (predominant reactivity with γδ T cells) CACT77A IgM nt nt nt nt nt Activation Molecule 8 (B cells) CACT65A IgM nt nt nt nt nt nt nt nt SLA Class I (Porcine MHC Class I) IgG2b nt nt nt +* nt nt nt nt nt nt nt nt Granulocytes PG68A IgG nt nt nt nt nt nt nt nt Granulocytes CL35A IgG1 nt nt nt nt nt + nt nt nt nt nt nt Granulocytes RACT43A IgM nt nt nt nt nt nt nt nt nt nt nt + Granulocytes + Endothelium GS23C IgG nt nt Granulocytes + Endothelium (G1) CH138A IgM nt nt Granulocytes + Endothelium (G1 cluster) MM20A IgG nt nt nt nt nt nt nt nt nt Granulocyte + Lymphocyte subpopulation E48C IgM nt nt nt nt + nt nt nt nt nt nt nt Granulocyte, Monocyte, Lymphocyte subpopulation CAT36A IgM nt nt nt nt nt + nt nt nt nt nt nt Granulocyte, Monocyte, Lymphocyte subpopulation PGBL1A IgM nt nt nt + nt nt nt nt nt nt nt nt Granulocyte, Monocyte, Lymphocyte subpopulation PGBL18A IgM nt nt nt + nt nt nt nt nt nt nt nt Platelets (85 kd) GB20A IgG nt nt nt nt nt nt nt nt Pan Leukocytes (CD45-like); non-reactive with platelets 25-2C IgM nt nt nt nt nt + nt nt nt nt nt nt Pan Leukocytes RACT38A IgG1 nt nt nt nt nt nt nt nt nt nt nt + Pan Leukocytes CF4A IgG nt - nt nt Pan Leukocytes CF11A IgM nt - nt nt Pan Leukocytes RT23A IgM nt nt nt nt nt nt nt nt nt nt nt + 38 * polymorphic

39 Monoclonal Antibodies to Immunoglobulins Specificity Cell Line Isotype Cattle Goat Sheep Swine Horse Cat Dog Ferret Mink Human Mouse Rabbit Bovine IgA BIG312D3 IgG1 + nt nt nt nt nt nt nt Bovine IgG1 BIG715A IgG nt nt nt nt Bovine IgM BIG73A IgG nt nt nt nt Bovine Ig λ Light Chain BIG501E IgG nt nt nt nt Equine IgM 1.9/3.2 IgG nt nt nt nt nt nt nt Porcine IgM PG145A IgM nt nt nt + nt nt nt nt nt nt nt nt Porcine IgM PIG45A IgG2b nt nt Monoclonal Antibodies to MHC Complex ANTIBODIES Specificity Cell Line Isotype Cattle Goat Sheep Swine Horse Cat Dog Ferret Mink Human Mouse Rabbit MHC Class I B5C IgG2b nt nt nt nt + - nt MHC Class I H17A IgG2a/2b +* - +/-* +* - nt nt nt nt +* - nt MHC Class I H58A IgG2a + +* + +* + +* MHC Class I PT85A IgG2a nt nt nt + - nt MHC Class I CF298A IgG1 nt nt nt nt nt + nt nt nt nt nt nt MHC Class II CAT82A IgG nt nt nt nt nt nt MHC Class II EqT2 IgG1 nt nt nt nt + nt nt nt nt nt nt nt MHC Class II MSA3 IgG2a nt nt nt nt nt - nt MHC Class II TH16B IgG2a * nt - nt MHC Class II TH21A IgG2b nt nt nt nt +* - nt MHC Class II (HLA-DP?) H42A IgG2a * nt MHC Class II (HLA-DQ) TH22A5 IgG2a nt nt nt nt nt - nt MHC Class II (HLA-DQ) TH81A5 IgG2a * * +* +* MHC Class II (HLA-DQ?) BAQ150A IgG nt + nt nt nt nt nt nt nt MHC Class II (HLA-DR) H34A IgG2b * nt MHC Class II (HLA-DRα) TH14B IgG2a * Monoclonal Antibodies to Cytokines and Hormones Specificity Cell Line Isotype Cattle Goat Sheep Swine Horse Cat Dog Ferret Mink Human Mouse Rabbit Bovine Interleukin-1β (IL-1β) IL IgG1 + nt nt nt nt nt nt nt nt nt nt nt Bovine Interleukin-1β (IL-1β) IL IgG1 + nt nt nt nt nt nt nt nt nt nt nt Bovine Interleukin-2 (reacts only with denatured IL-2) IL IgG1 + nt nt nt nt nt nt nt nt nt nt nt Bovine Interleukin-2 (reacts only with denatured IL-2) IL IgG1 + nt nt nt nt nt nt nt nt nt nt nt Bovine Granulocyte-Macrophage Colony-Stimulating Factor GM-CSF 17.2 IgG1 + nt nt nt nt nt nt nt nt nt nt nt Bovine Granulocyte-Macrophage Colony-Stimulating Factor GM-CSF 20.1 IgG1 + nt nt nt nt nt nt nt nt nt nt nt Bovine Growth Hormone B24.3 IgM + nt nt nt nt nt nt nt nt nt nt nt Bovine Growth Hormone B8.10 IgG1 + nt nt nt nt nt nt nt nt nt nt nt Bovine Somatostatin SS1.1 NT + nt nt nt nt nt nt nt nt nt nt nt Porcine Growth Hormone PS8.3 IgG2a nt nt nt + nt nt nt nt nt nt nt nt 39 * polymorphic

40 Coombs Reagents The Coombs test, also called direct antiglobulin test, is designed to detect immune-mediated erythrocyte destruction which occurs in autoimmune hemolytic anemias, and in some cases with infections and neoplastic disorders. Hemolysis in these diseases is caused by the erythrocytes being coated with antibody (IgG, IgM) and/or complement components (C3). Coated erythrocytes are either lysed in the bloodstream or removed by phagocytes. VMRD s Coombs reagent is a caprine-origin antiserum against IgG, IgM, and C3. It does not agglutinate normal erythrocytes, but does agglutinate erythrocytes coated with IgG, IgM, and/or C3. Agglutination, which may be observed macroscopically or microscopically, is indicative of a Coombs positive. Canine Coombs All dogs with anemia (including that caused by intravascular and extravascular hemolysis) of unknown origin are reasonable candidates for evaluation by Coombs testing. VMRD s Canine Anti-Sheep Red Blood Cells (SRBC) are used to prepare positive controls. Canine Coombs Test Size Catalog No. Canine Coombs 2 ml ml Canine Anti-Sheep Red Blood Cells (SRBC) 1 ml Equine Coombs All horses with anemia (including that caused by intravascular and extravascular hemolysis) of unknown origin are reasonable candidates for evaluation by Coombs testing. Foals with neonatal isoerythrolysis are often Coombs positive. Equine Coombs Test Size Catalog No. Equine Coombs 2 ml IMMUNOLOGY Feline Coombs All cats with anemia (including that caused by intravascular and extravascular hemolysis) of unknown origin are reasonable candidates for evaluation by Coombs testing. Feline Coombs Test Size Catalog No. Feline Coombs 2 ml

41 Radial Immunodiffusion (RID) Kits VMRD s Single Radial Immunodiffusion (SRID) kits offer a convenient and accurate means of quantification of immunoglobulin levels in serum, plasma, and colostrum. Each kit contains pre-poured plates and four standards of known immunoglobulin concentration. Using the kit is as simple as adding the standards and samples to be tested to plate wells and incubating the plate overnight at room temperature using the procedure as established by Fahey. Monospecific anti-immunoglobulin antiserum contained in the agarose gel on the plate binds immunoglobulin in standards and samples forming sharp, easy-to-read precipitin rings of corresponding diameter to immunoglobulin concentration. Ring sizes and immunoglobulin concentrations of the four standards are used to develop a standard curve from which sample immunoglobulin concentrations may be calculated. A semi-log coordinate system included on the package insert straightens the standard curve to improve the accuracy of interpolation of sample concentrations. No complex calculations are necessary; the investigator simply finds the point on the standard curve where the X coordinate matches the sample ring diameter and then reads the immunoglobulin concentration from the corresponding Y coordinate. Test Specificity Isotype Range (mg/100 ml) Size Catalog No. Bovine IgA determinations determinations IgG determinations determinations IgG determinations determinations IgG determinations determinations IgM determinations determinations Canine IgA determinations determinations IgG determinations determinations IgM determinations determinations IMMUNOLOGY Equine IgA determinations determinations IgG determinations determinations IgM determinations determinations IgG(T) determinations determinations Feline IgG determinations determinations Porcine IgG determinations determinations

42 Failure of Passive Transfer (FPT) Field Tests FPT Field Test Size Catalog No. 10 tests BOVA-S 24 tests BOVA-S A Field Test for Diagnosis of Failure of Passive Transfer in Calves Failure of passive transfer (FPT) is the underlying cause of most neonatal calf losses associated with gastroenteritis, pneumonia, septicemia, omphalophlebitis, and polyarthritides. Prevalence of FPT varies with management, conditions, and type of animal (beef or dairy), but is usually greater than 10% and may be as high as 60% in some dairy herds. Depending on the pathogens and conditions faced, risk of death among calves with FPT is 3 to 10 times greater than among calves that absorb adequate amounts of immunoglobulins. Management practices that increase absorption of colostral antibodies clearly decrease calf losses and increase profits. One key to preventing losses from FPT is a fast and accurate diagnosis. Whether you are spot testing a commercial herd or testing every pure-bred calf, you do not have time to wait for lab results. You need a convenient and reliable field test BOVA-S. BOVA-S gives you a definitive diagnosis of the FPT status of the calf in 30 minutes. All you do is add serum. FPT Field Test Size Catalog No. 5 tests EQUI-Z 10 tests tests EQUI-Z A Field Test for Diagnosis of Complete and Partial Failure of Passive Transfer in Equine Foals The most common conditions leading to infectious diseases in neonatal foals are failure of passive transfer (FPT) and partial failure of passive transfer (PFPT) of colostral immunoglobulins from mare to foal. Even wellmanaged brood farms can expect FPT (less than 200 mg IgG/100 ml of serum) or PFPT ( mg IgG/100 ml of serum) in 20-25% of their foals due to factors largely beyond their control. Up to 75% of foals with FPT and 25% of foals with PFPT develop infectious disease and nearly all foals that die of infectious disease have less than 400 mg IgG/100 ml of serum. Therefore, screening of foals for failure of passive transfer should be a routine part of neonatal health care. With EQUI-Z you do not have to waste time waiting for lab results. EQUI-Z can determine the FPT or PFTP status of your foal in the field in just one hour. All you do is add serum. IMMUNOLOGY 42 FPT Field Test Size Catalog No. LLAMA-S 5 tests LLAMA-S A Field Test for Diagnosis of Failure of Passive Transfer in Llamas Failure of passive transfer (FPT) of colostral immunoglobulins is a major determinant of infections and mortality in newborn llamas. Protective immunoglobulins are passed from the dam through colostrum and absorbed by the cria. LLAMA-S is a convenient field test for adequate passive transfer of these colostral antibodies to the newborn llama. All you do is add serum. LLAMA-S also works with Alpacas.

43 Ordering Information Orders may be placed by , FAX, telephone or mail/post. FAX: Phone: Mail: VMRD, Inc.. P.O. Box 502. Pullman, WA USA Business Hours: Backorders: Custom Orders: Returns: Monday-Friday, 7 am - 5 pm (Pacific Time Zone) Out-of-stock items are placed on backorder and shipped as soon as available. Custom orders are prepared on a contract basis only. Please contact us for information. Call for authorization prior to returning any item. Returns are subject to a 25% restock fee. Custom orders may not be returned. Technical Assistance: Our staff is available to assist as needed. Consulting and research services are available on a contract basis. Product Information: For information throughout the year on VMRD products visit our website, send an to [email protected] or [email protected]; or call Warranty: VMRD, Inc. warrants that the products are as described as to the quantity and contents stated on the label at the time of delivery to the customer. NO OTHER WARRANTIES, EXPRESS OR IMPLIED, ARE MADE BEYOND THE LABEL DESCRIPTION, INCLUDING WARRANTIES OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR USE. Remedy is limited to replacement of the product or refund of the purchase price. VMRD, Inc. is not liable for property damage, personal injury, or economic loss caused by the product. Ordering Procedures When placing an order, please supply the appropriate customer identification number, catalog number(s) and quantity of the items needed, and a brief description of each product. Invoicing Procedures Billing invoices are mailed separately following shipment. Payment terms are Net 30 days, payable in U.S. Dollars. Please inquire to arrange payment by wire transfer. Payment may also be made by Visa, MasterCard, or American Express credit cards. Please specify payment by credit card when the order is placed. Invoice questions may be directed to our Customer Service Department at Shipping Procedures Most items ship within one business day from the date the order was received, except where special certificates are required. Shipping fees are prepaid and added to the invoice, unless the recipient provides courier account information. International Orders International orders should include a copy of any necessary import permits or other documentation required for customs clearance. Payment of duties and taxes are the responsibility of the recipient. Requirement of an Export Certificate Animal Products signed by a United States federal veterinarian should be indicated at the time an order is placed. The cost to obtain said certificate from VMRD is currently $65 USD. Volume Discounts VMRD, Inc. offers volume discounts on test kits. Other items in the catalog are also subject to volume discount. Please inquire with the Marketing Department at VMRD at [email protected] or FAX: Toll Free in the USA and Canada: [email protected]

44 VMRD Distributors VMRD Distributors VMRD has distributors in many countries, including Argentina, Australia, Belgium, Bosnia and Herzegovina, Brazil, Bulgaria, Canada, Chile, China, Costa Rica, Cyprus, Czech Republic, Dominican Republic, Ecuador, El Salvador, France, Germany, Greece, Guatemala, Honduras, Hong Kong, Hungary, India, Iraq, Ireland, Israel, Italy, Japan, Mexico, New Zealand, Nicaragua, Peru, Philippines, Poland, Romania, Saudi Arabia, Serbia & Montenegro, Slovenia, South Korea, Spain, Switzerland, Taiwan, Turkey, Ukraine, The United Arab Emirates and Venezuela. Why We Use Distributors Distributors help our customers by overcoming limitations imposed by distance, language, culture, and trade restrictions. Furthermore, they bring our products to customers who would not otherwise know about them. They also save our customers money by importing in quantity at discounted prices. In addition, they provide first-level technical support and serve as a liaison for second-level technical support. How to Find a Distributor Please visit our website ( for a current list of distributors, or give us a call ( ) or ([email protected]) to inquire whether we have a distributor in your country. Photo Courtesy Mansfeild Jones 44

45 VMRD, Inc Pullman-Albion Road Pullman, WA USA Phone: Fax: International Fax/Mail Order Form Please Photocopy for Future Use Order Date: Shipping Address Company Address Customer ID: Billing Address Company Address City Postal Code Attention Phone Fax Order Details Purchase Order No. State Country Credit Card No. SPECIAL INSTRUCTIONS City Postal Code Attention Phone Fax State Country Exp. Date Catalog Number Quantity Description Unit Price Total Price Document Preparation Charge $10.00 $10.00 *TOTAL (USD): * Shipping Charges will be prepaid and added to the invoice total. Orders are shipped D.D.U. (Delivery Duty Unpaid). Payment Options: Wire Transfer, VISA/MC or American Express credit cards, Check in United States Dollars

46 Index by Catalog Number Catalog No. Page AP AP AP CDV CDV REO REO CDV CDV NC NC CHV CHV CC CC CN CN CSO ERV ERV CS CS PCRV RAB RAB PCRV FIP...27 Catalog No. Page CPI CPI CAV CAV BT BT FPL PAV PAV FPL FELV FELV FELV FVR TGE PPV PI PI BAV TGE BAV BAV BAV PPV BVD BVD BRSV BRSV BCV BCV BPV BPV IBR IBR BAV BRSV BVD CCV CDV ERV FIP FIP FPL IBR NC PCRV PI PPV TGE TOXO VSV...28 Catalog No. Page BLV BRSV BT BVD CAV CCV CDV CHV CPI CPV ERV FCV FIP FIP FPL FVR IBR PCRV PI PPV TGE BBI BBO BH C CB CC CN CS CSO EC EE FC LD LSH NC RMSF TOXO BLV BRSV BVD CAV CCV CDV CPI CPV ERV FCV FELV FIP FIP Catalog No. Page FIP FPL FVR IBR PCRV PI PPV RAB TGE RB CB MF SB SB N-BB N-BBI N-BLV N-BRSV N-BTV N-BVD N-CAV N-CB N-CPI N-EC N-EE N-FCV N-FIP N-FIP N-IBR N-LD N-LSH N-NC-BOV N-NC-CAN N-PCRV N-PI N-RMSF N-TGE P-BB P-BBI P-BH-G P-BH-M P-BLV P-BRSV P-BTV P-BVD P-CAV P-CDV-G P-CDV-M P-CHV P-CPI P-CPV-G...25

47 Catalog No. Page 211-P-CPV-M P-EC P-EE P-ERV-EQ P-ERV-LL P-FCV P-FHV P-FIP P-FIP P-IBR P-LD P-LSH P-NC-BOV P-NC-CAN P-PCRV P-PI P-RMSF P-TGE P-TOXO-FEL N-CDV N-FELV P-CDV P-FELV ELISAWARE... 5

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